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A Review of the Environmental Safety of the PAT ProteinCenter for Environmental Risk Assessment, ILSI Research Foundation1156 Fifteenth Street N.W., Washington D.C. 20005-1743 USA
April 11, 2011
Key words
PAT, bar, phosphinothricin, glu-fosinate, glufosinate ammonium, herbicide tolerant, genetically engineered, environmental risk assessment
Copyright © ILSI Research Foundation 2011This work is licensed under the Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 United States License. To view a copy of this license, visit http://creativecommons.org/l icenses/by-nc-nd/3.0/us/ or send a letter to Creative Commons, 171 Second Street, Suite 300, San Francisco, California, 94105, USA.
IntroductIon
This document provides a comprehensive review of information and data relevant to the environ-mental risk assessment of the protein phosphi-nothricin-N-acetyl transferase (PAT) produced in genetically engineered (GE) plants by genes isolated from Streptomyces viridochromogenes (pat gene) or Streptomyces hygroscopicus (bar gene) and presents a summary statement about the environ-mental safety of this protein. All sources of infor-mation reviewed herein were publicly available and include: dossiers presented to regulatory authori-ties; decision summaries prepared by regulatory authorities; peer-reviewed literature; and prod-uct summaries prepared by product developers. Many GE plants contain the pat gene for use as a selectable marker during development. In those cases, there are one or more additional transgenes contained in the plant and the final product is not necessarily glufosinate tolerant. Although this doc-ument will not address these additional genes and phenotypes, their presence should be noted when looking at data on the GE plants that express PAT.
Environmental risk assessments related to the in-troduction of GE plants are conducted on a case-by-case basis taking into account the biology of the plant, the nature of the transgene and the protein or gene product it produces, the phenotype con-ferred by the transgene, as well as the intended use of the plant and the environment where it will be introduced (i.e., the receiving environment). These assessments typically involve comparisons of the transgenic event to an untransformed parent line and/or closely related isoline, and also use base-line knowledge of the relevant plant species (CBD 2000b, Codex 2003a, b, EFSA 2006a, NRC 1989, OECD 1992, OECD 2006). The objective of these comparisons is to identify potential risks that the GE plant might present beyond what is already accepted for similar plants in the environment by identifying meaningful differences between the GE crop and its conventional counterpart. Any identified differences that have the potential to
cause relevant adverse effects can subsequently be evaluated for likelihood and consequence.
To date, regulatory authorities in 11 different countries have issued approvals for the environ-mental release of GE plants expressing the PAT protein, either by itself or in combination with other GE traits. This represents approximately 38 transformation events and includes 8 species of plant: Beta vulgaris L. (sugarbeet), Brassica napus L. and Brassica rapa L. (oilseed rape and turnip rape, respectively, although both can be re-ferred to as canola), Cichorium intybus (chicory), Glycine max L. (soybean), Gossypium hirsutum L. (cotton), Oryza sativa L. (rice) and Zea mays L. (maize). These regulatory analyses have generally considered three categories of potential harm: (1) the PAT protein may have an adverse impact on non-target organisms; (2) transformation of the host plant and subsequent expression of the PAT protein may alter the characteristics of the plant, resulting in adverse environmental impacts (e.g. increased weediness); and (3) introgression of the gene encoding the PAT protein into a sexu-ally compatible plant species may alter that species resulting in adverse environmental impacts (e.g. establishment of new weedy populations) (CFIA 1995a, 1995b, 1996a, 1996b, 1996c, 1996d, 1996e, 1996f, 1998a, 1998b, 1998c, 1999, 2002a, 2002b, 2005, 2006, EC 1996, 1997, 1998, 2001, Japan BCH 1996a, 1996b, 1996c, 1997a, 1997b, 1997c, 1997d, 1997e, 1998, 1999a, 1999b, 1999c, 1999d, 2002, 2005, 2006a, 2006b, 2006c, 2006d, 2006e, 2007a, 2007b, 2008, 2009, 2010, OGTR 2002, 2003, 2006, Philippines 2005, USDA APHIS 1995a, 1995c, 1995f, 1996b, 1996b, 1996c, 1996e, 1997c, 1997f, 1998a, 1998c, 1998e, 1998g, 1998i, 1998j, 1999a, 1999b, 1999c, 2001b, 2001c, 2002b, 2003c, 2004a, 2004c, 2005, 2006b, USEPA 2001, 2005, 2009a, 2009b).
Note that environmental effects that may be as-sociated with the use of the herbicide glufosinate in association with GE plants producing PAT are outside the purview of this review.
2
Tabl
e 1.
Reg
ulat
ory
appr
oval
s for
the
envi
ronm
enta
l rel
ease
of G
E pl
ants
cont
aini
ng P
AT p
rote
in.
Spec
ies
Even
ts/C
ross
esA
lter
nate
Des
igna
tion
sSo
urce
of P
AT G
ene
United States
Canada
Japan
Australia
European Union
Brazil
Argentina
Colombia
Philippines
South Africa
Uruguay
Beta
Vul
garis
(sug
arbe
et)
ACS-
BVØ
Ø1-
3T
120-
7St
rept
omyc
es vi
ridoc
hrom
ogen
esX
X
Bras
sica
napu
s (oi
lseed
rape
/can
ola)
HC
N1Ø
Stre
ptom
yces
virid
ochr
omog
enes
XX
X
ACS-
BNØ
Ø7-
1H
CN
92St
rept
omyc
es vi
ridoc
hrom
ogen
esX
XX
X
ACS-
BNØ
Ø4-
7 x
ACS-
BNØ
Ø1-
4M
S1, R
F1 ;
PGS1
Stre
ptom
yces
hygr
osco
picu
sX
XX
X
ACS-
BNØ
Ø4-
7 x
ACS-
BNØ
Ø2-
5M
S1, R
F2 ;
PGS2
Stre
ptom
yces
hygr
osco
picu
sX
XX
X
ACS-
BNØ
Ø5-
8 x
ACS-
BNØ
Ø3-
6M
S8 x
RF3
Stre
ptom
yces
hygr
osco
picu
sX
XX
X
PHY1
4, P
HY3
5St
rept
omyc
es hy
gros
copi
cus
X
PHY3
6St
rept
omyc
es hy
gros
copi
cus
X
ACS-
BNØ
Ø8-
2T
45, H
CN
28St
rept
omyc
es vi
ridoc
hrom
ogen
esX
XX
X
Bras
sica
rapa
(bird
rape
/can
ola)
HC
R-1
Stre
ptom
yces
virid
ochr
omog
enes
X
Cich
oriu
m in
tybu
s (ch
icor
y)R
M3-
3, R
M3-
4, R
M3-
6St
rept
omyc
es hy
gros
copi
cus
XX
Glyc
ine m
ax (s
oybe
an)
ACS-
GM
ØØ
5-3
A270
4-12
, A27
04-2
1, A
5547
-35
Stre
ptom
yces
virid
ochr
omog
enes
XX
XX
ACS-
GM
ØØ
6-4
A554
7-12
7St
rept
omyc
es vi
ridoc
hrom
ogen
esX
XX
X
ACS-
GM
ØØ
3-1
GU
262
Stre
ptom
yces
virid
ochr
omog
enes
X
ACS-
GM
ØØ
1-8,
AC
S-G
MØ
Ø2-
9W
62, W
98St
rept
omyc
es hy
gros
copi
cus
X
Gos
sypi
um h
irsut
um (c
otto
n)D
AS-2
4236
-528
1-24
-236
Stre
ptom
yces
virid
ochr
omog
enes
X
DAS
21Ø
23-5
3006
-210
-23
Stre
ptom
yces
virid
ochr
omog
enes
X
DAS
21Ø
23-5
x D
AS-2
4236
-5St
rept
omyc
es vi
ridoc
hrom
ogen
esX
X
DAS
21Ø
23-5
x D
AS-2
4236
-5 x
MO
N-Ø
1445
-2St
rept
omyc
es vi
ridoc
hrom
ogen
es*
DAS
21Ø
23-5
x D
AS-2
4236
-5 x
MO
N-8
8913
-8St
rept
omyc
es vi
ridoc
hrom
ogen
es*
ACS-
GH
ØØ
1-3
LLC
otto
n25
Stre
ptom
yces
hygr
osco
picu
sX
XX
ACS-
GH
ØØ
1-3
x M
ON
-159
85-7
LLC
otto
n25
x M
ON
1598
5St
rept
omyc
es hy
gros
copi
cus
*X
Ory
za sa
tiva
(ric
e)AC
S-O
SØØ
1-4,
AC
S-O
SØØ
2-5
LLR
ice0
6, L
LRic
e62
Stre
ptom
yces
hygr
osco
picu
sX
BCS-
OSØ
Ø3-
7LL
Ric
e601
Stre
ptom
yces
hygr
osco
picu
sX
Zea
may
s (m
aize
/cor
n)SY
N-E
V17
6-9
176
Stre
ptom
yces
hygr
osco
picu
sX
XX
XX
PH-Ø
ØØ
676-
7, P
H-Ø
ØØ
678-
9, P
H-Ø
ØØ
68Ø
-267
6, 6
78, 6
80St
rept
omyc
es vi
ridoc
hrom
ogen
esX
DK
B-89
79Ø
-5B1
6, D
LL25
Stre
ptom
yces
hygr
osco
picu
sX
XX
SYN
-BTØ
11-1
BT11
(X43
34C
BR, X
4734
CBR
)St
rept
omyc
es vi
ridoc
hrom
ogen
esX
XX
XX
XX
XX
SYN
-BTØ
11-1
x M
ON
-ØØ
Ø21
-9BT
11 x
GA2
1St
rept
omyc
es vi
ridoc
hrom
ogen
es*
XX
X
SYN
-BTØ
11-1
x S
YN-I
R16
2-4
BT11
x M
IR16
2St
rept
omyc
es vi
ridoc
hrom
ogen
esX
SYN
-BTØ
11-1
x S
YN-I
R16
2-4
x SY
N-I
R6Ø
4-5
BT11
x M
IR16
2 x
MIR
604
Stre
ptom
yces
virid
ochr
omog
enes
X
SYN
-BTØ
11-1
x S
YN-I
R6Ø
4-5
BT11
x M
IR60
4St
rept
omyc
es vi
ridoc
hrom
ogen
es*
X
SYN
-BTØ
11-1
x SY
N-IR
6Ø4-
5 x M
ON
ØØ
Ø21
-9BT
11 x
MIR
604
x G
A21
Stre
ptom
yces
virid
ochr
omog
enes
*
ACS-
ZM
ØØ
4-3
CBH
-351
Stre
ptom
yces
hygr
osco
picu
sX
DAS
-Ø62
75-8
Stre
ptom
yces
hygr
osco
picu
sX
XX
DAS
-591
22-7
Stre
ptom
yces
virid
ochr
omog
enes
XX
X
DAS
-591
22-7
, MO
N-Ø
Ø6Ø
3-6
DAS
-591
22-7
x N
K60
3St
rept
omyc
es vi
ridoc
hrom
ogen
es*
XX
Tabl
e 1
(con
tinu
ed o
n ne
xt p
age)
.
3
orIgIn and FunctIon oF Pat
Phosphinothricin, bialaphos, and glufosinate ammonium
In the early 1970’s a previously unknown amino acid was isolated independently from two species of Streptomyces by laboratories working in Germany (from Streptomyces viridochromogenes) and Japan (from Streptomyces hygroscopicus) (Bayer et al. 1972, Kondo et al. 1973, OECD 1999). Originally seen in a tripeptide with two alanine residues (see Fig. 1), the new amino acid (L-2-amino-4-[hydroxyl(methyl)phosphinyl] butyric acid) was given the name phosphinothricin (PT) and the tripeptide called phosphinothricin tripeptide (PTT) or bialaphos1 (Bayer et al. 1972, Hoerlein 1994, Kondo et al. 1973, OECD 1999). In Germany, racemic mixtures were produced (D,L-phosphinothricin or D,L-PPT) and deter-mined to have herbicidal activity. D,L-PPT-ammonium, referred to by the common name glufosinate ammonium (GLA) is the active ingredient in herbicide formulations marketed worldwide. In Japan, the bialaphos tripeptide was observed to have herbicidal activity and this has been commercialized as well (Hoerlein 1994).
Figure 1. The structure of phosphinothricin, PTT and glutamic acid.
CH3
O ═ P ─ OH
CH2
CH2
H ─ C ─ NH2
O ═ C ─ Ala ── Ala
phosphinothricin tripeptide(PTT)
CH3
O ═ P ─ OH
CH2
CH2
H ─ C ─ NH2
O ═ C ─ OH
phosphinothricin(PT)
O ═ C ─ OH
CH2
CH2
H ─ C ─ NH2
O ═ C ─ OH
glutamic acid 2
Phosphinothricin inhibits the activity of the glutamine synthetase enzyme (GS) by competitively binding in place of the normal sub-strate, glutamate (glutamic acid). This prevents the synthesis of L-glutamine, which is not only an important chemical precursor for the synthesis of nucleic acids and proteins, but serves as the mechanism of ammonia (NH3) incorporation for plants (Hoerlein 1994, OECD 1999, OECD 2002). Treatment with phosphi-nothricin causes accumulation of ammonia and cessation of pho-tosynthesis, probably due to the lack of glutamine (Hoerlein 1994, OECD 1999, OECD 2002).
IsolatIon and FunctIon oF PhosPhInothrIcIn acetyl transFerase (Pat)
The identification of the GS inhibitor phosphinothricin from Streptomyces suggested that these bacteria employ a biochemical mechanism to preserve endogenous GS activity. In the late 1980s,
1 Also sometimes “bilanafos” or “bilanaphos”.
2 Figure adapted from Schwartz et al. 2004.Spec
ies
Even
ts/C
ross
esA
lter
nate
Des
igna
tion
sSo
urce
of P
AT G
ene
United States
Canada
Japan
Australia
European Union
Brazil
Argentina
Colombia
Philippines
South Africa
Uruguay
DAS
-591
22-7
x D
AS-Ø
15Ø
7-1
x MO
N-Ø
Ø6Ø
3-6
DAS
-591
22-7
x T
C15
07 x
NK
603
Stre
ptom
yces
virid
ochr
omog
enes
*X
X
DK
B-89
614-
9D
BT41
8St
rept
omyc
es hy
gros
copi
cus
XX
XX
MO
N-8
9Ø34
-3 x
DAS
- Ø15
Ø7-
1 x
MO
N-
88Ø
17-3
x D
AS-5
9122
-7M
ON
8903
4 x
TC
1507
x
MO
N88
017
x D
AS-5
9122
-7St
rept
omyc
es vi
ridoc
hrom
ogen
esX
XX
ACS-
ZM
ØØ
1-9
MS3
Stre
ptom
yces
hygr
osco
picu
sX
X
ACS-
ZM
ØØ
5-4
MS6
Stre
ptom
yces
hygr
osco
picu
sX
MO
N-Ø
Ø6Ø
3-6
x AC
S-Z
MØ
Ø3-
2N
K60
3 x
T25
Stre
ptom
yces
virid
ochr
omog
enes
*X
ACS-
ZM
ØØ
2-1,
AC
S-Z
MØ
Ø3-
2T
14, T
25St
rept
omyc
es vi
ridoc
hrom
ogen
esX
XX
XX
X
ACS-
ZM
ØØ
3-2,
MO
N-Ø
Ø81
Ø-6
T25
x M
ON
810
Stre
ptom
yces
virid
ochr
omog
enes
*X
DAS
-Ø15
Ø7-
1T
C15
07St
rept
omyc
es vi
ridoc
hrom
ogen
esX
XX
XX
DAS
-Ø15
Ø7-
1, D
AS-5
9122
-7T
C15
07 x
DAS
-591
22-7
Stre
ptom
yces
virid
ochr
omog
enes
*X
X
DAS
-Ø15
Ø7-
1 x
MO
N-Ø
Ø6Ø
3-6
TC
1507
x N
K60
3St
rept
omyc
es vi
ridoc
hrom
ogen
es*
XX
XX
X =
App
rove
d fo
r env
ironm
enta
l (co
mm
erci
al) r
elea
se.
* Th
ese
are
stack
ed e
vent
s tha
t may
be
cons
ider
ed a
ppro
ved
in th
e co
untr
y in
dica
ted
Tabl
e 1
(con
t’d).
Regu
lato
ry a
ppro
vals
for t
he e
nviro
nmen
tal r
elea
se o
f GE
plan
ts co
ntai
ning
PAT
pro
tein
.
4
two genes were identified independently based on their ability to confer resistance to phosphinothricin inhibition of GS, both of which encode a phosphinothricin acetyl transferase protein (PAT). The bialaphos resistance gene, bar, was isolated from S. hygroscopicus while the homologous gene from S. viridochromogenes was termed pat after the function of the enzyme (OECD 1999a, Thompson et al. 1987, Wohlleben et al. 1998). Both proteins have been used extensively in genetic engineering of crop plants. They both con-sist of 183 amino acids, with a sequence identity of 85% (OECD 1999a, Wehrmann et al. 1996, Wohlleben et al. 1998). Importantly, both proteins acetylate phosphinothricin but show no activity with glutamate, which is structurally similar, or with any other amino acids tested, indicating a high specificity (OECD 1999a, Thompson et al. 1987, Wehrmann et al. 1996). The only recorded differences in activity between the two proteins are minor differences in the optimal pH, and a significantly different affinity for acetyl-coA (a co-substrate); these differences are not expected to be meaningful in planta (OECD 1999a, Wehrmann et al. 1996). Because the PAT proteins en-coded by bar and pat are structurally and functionally equivalent, with similar molecular weights, immuno-cross-reactivity, substrate affinity and specificity, they are considered together in this document and will both be referred to as PAT protein.
The PAT enzyme acetylates phosphinothricin at the N-terminus. N-acetyl phosphinothricin has no her-bicidal activity, and resistance is therefore conferred through modification of the herbicide rather than the target of its activity (OECD 1999a, Thompson et al. 1987, Wehrmann et al. 1996, Wohlleben et al. 1998).
exPressIon oF Pat In PhosPhInothrIcIn-tolerant ge Plants
Data for the level of expression of PAT in phosphinothricin-toler-ant GE plants that have obtained regulatory approvals are available in publicly accessible regulatory documents (ANZFA 2000, 2001a, 2001b, 2002, CFIA 1995a, 1995b, 1996a, 1996b, 1996c, 1996d, 1996e, 1996f, 1998a, 1998b, 1998c, 1999, 2002a, 2002b, 2004, 2005, 2006a, 2006b, EC 1996, 1997, 1998, 2001, EFSA 2005, 2006, 2008a, 2008b, 2009a, 2009b, FSANZ 2003, 2004a, 2004b, 2005a, 2005b, 2008, Japan BCH 1996a, 1996b, 1996c, Health Canada 2006a, 2006b, Japan BCH 1997a, 1997b, 1997c, 1997d, 1997e, 1999a, 1999b, 1999c, 1999d, 2002, 2003, 2005, 2006a, 2006b, 2006c, 2006d, 2006e, 2006f, 2007a, 2007b, 2008, 2009, 2010, OGTR 2003, 2006, Philippines 2005, USDA APHIS 1994a, b, 1995b, 1995d, 1995e, 1996a, 1996d, 1997a, 1997b, 1997d, 1997e, 1998b, 1998d, 1998f, 1998h, 1998k, 1998l, 2000, 2001a, 2002a, 2002c, 2003a, 2003b, 2004b, 2004d, 2006a, USEPA 2001, 2005, 2009a, 2009b). Tissue types tested and sampling methodol-ogies vary greatly. The most common method uses enzyme-linked immunosorbent assay (ELISA) to quantify the amount of protein present in a given sample, but other methods include an assay for enzymatic activity and the use of Northern blots to quantify mRNA. Normally, one or more samples are collected from plants
in field trials or greenhouse experiments and the amount of protein is given as a mean accompanied by either a standard deviation or a range of observed values to show variability. The result is often quantified as a ratio to the dry weight of the sample (e.g. µg PAT/g dry weight), but some reports calculate the ratio to the fresh weight of the sample or to the total extractable protein from the sample (e.g. µg PAT/g total protein).
Variations in methodology for both sample collection and subse-quent analysis make direct statistical comparisons of the data inap-propriate. However, the weight of evidence suggests PAT protein is expressed at low levels (see Annex I and associated references). The highest reported levels of expression observed in each species using ELISA are reported in Table 2.
establIshment and PersIstence oF Pat-exPressIng Plants In the envIronment
Familiarity with the biology of the non-transformed or host plant species in the receiving environment is typically the starting point for a comparative environmental risk assessment of a GE plant (CBD 2000b, Codex 2003a, b, EFSA 2006a, NRC 1989, OECD 1992, OECD 2006). Information about the biology of the host plant can be used to identify species-specific characteristics that may be affected by the novel trait so as to permit the transgenic plant to become “weedy,” invasive of natural habitats, or to be oth-erwise harmful to the environment. It can also provide details on significant interactions between the plant and other organisms that may be important when considering potential harms. By consider-ing the biology of the host plant, a risk assessor can identify poten-tial hazards that may be associated with the expression of the novel protein (e.g., PAT) and then be able to assess the likelihood of these hazards being realized. For example, if the plant species is highly domesticated and requires significant human intervention to grow or reproduce, the assessor can take that into account when assessing the likelihood of the GE plant establishing outside of cultivation.
Table 2. Highest reported expression level of PAT protein using ELISA.1
Species Event GE Plant Expression Tissue Reference
Beta vulgaris T120-7 966 ng/g Top2 USDA APHIS 1998b
Brassica napus Topas19/2 x T45 944 ng/g Leaf USDA APHIS 2002a
Glycine max A5547-127 20202 ± 3593 ng/g
Seed Japan BCH 2005f
Gossypium hirsutum LLCOTTON25 127000 ± 180003 ng/g4
Cleaned Seed USDA APHIS 2002c
Oryza sativa LLRICE62 84700 ng/g4 Leaf CFIA 2006b
Zea mays DAS-06275-8 935000 ng/g4, 5 Leaf USDA APHIS 2004b
1 These values are not cross-comparable due to differences in sample collection and preparation methodology.2 Top refers to all above-ground tissue (i.e. leaves and stems).3 Reported as mean ± standard deviation.4 Reported as ng/g fresh weight.5 Represents the highest value in a reported range.
5
PhenotyPIc data
In order to determine if GE plants expressing PAT are phenotypi-cally different than their non-transformed counterparts, a variety of data have been collected and are presented with varying degree of detail in regulatory submissions related to the environmental release of these plants (CFIA 1995a, 1995b, 1996a, 1996b, 1996c, 1996d, 1996e, 1996f, 1998a, 1998b, 1998c, 1999, 2002a, 2002b, 2004, 2005, 2006a, 2006b, EC 1996, 1997, 1998, 2001, Japan BCH 1996a, 1996b, 1996c, Health Canada 2006a, 2006b, Japan BCH 1997a, 1997b, 1997c, 1997d, 1997e, 1999a, 1999b, 1999c, 1999d, 2002, 2003, 2005, 2006a, 2006b, 2006c, 2006d, 2006e, 2006f, 2007a, 2007b, 2008, 2009, 2010, OGTR 2003, 2006, Philippines 2005, USDA APHIS 1994a, b, 1995b, 1995d, 1995e, 1996a, 1996d, 1997a, 1997b, 1997d, 1997e, 1998b, 1998d, 1998f, 1998h, 1998k, 1998l, 2000, 2001a, 2002a, 2002c, 2003a, 2003b, 2004b, 2004d, 2006a, USEPA 2001, 2005, 2009a, 2009b). The data that are reported are dependent on the species of plant, but in general include information on gross morphology (e.g. height, number of leaves, number of branches or nodes, etc.), reproductive characteristics including seed production, survival and germina-tion, as well as seedling vigor, overwintering ability, susceptibility to disease and pest pressure, and frequently the potential to volunteer following harvest. Phenotypic analyses may also include agronomic characteristics such as yield and performance in the field (CFIA 1995a, 1995b, 1996a, 1996b, 1996c, 1996d, 1996e, 1996f, 1998a, 1998b, 1998c, 1999, 2002a, 2002b, 2004, 2005, 2006a, 2006b, EC 1996, 1997, 1998, 2001, Japan BCH 1996a, 1996b, 1996c, Health Canada 2006a, 2006b, Japan BCH 1997a, 1997b, 1997c, 1997d, 1997e, 1999a, 1999b, 1999c, 1999d, 2002, 2003, 2005, 2006a, 2006b, 2006c, 2006d, 2006e, 2006f, 2007a, 2007b, 2008, 2009, 2010, OGTR 2003, 2006, Philippines 2005, USDA APHIS 1994a, b, 1995b, 1995d, 1995e, 1996a, 1996d, 1997a, 1997b, 1997d, 1997e, 1998b, 1998d, 1998f, 1998h, 1998k, 1998l, 2000, 2001a, 2002a, 2002c, 2003a, 2003b, 2004b, 2004d, 2006a, USEPA 2001, 2005, 2009a, 2009b). Frequently, statistically significant differenc-es in a handful of phenotypic characteristics are reported between GE plants and controls in a given experiment (CFIA 1995a, 1995b, 1996a, 1996b, 1996c, 1996d, 1996e, 1996f,1998a, 1998b, 1998c, 1999, 2002a, 2002b, 2004, 2005, 2006a, 2006b, EC 1996, 1997, 1998, 2001, Japan BCH 1996a, 1996b, 1996c, Health Canada 2006a, 2006b, Philippines 2005, USDA APHIS 1994a, b, 1995b, 1995d, 1995e, 1996a, 1996d, 1997a, 1997b, 1997d, 1997e, 1998b, 1998d, 1998f, 1998h, 1998k, 1998l, 2000, 2001a, 2002a, 2002c, 2003a, 2003b, 2004b, 2004d, 2006a, USEPA 2001, 2005, 2009a, 2009b). However, these differences usually are not repeated in mul-tiple experiments and regulatory decisions have concluded that any such differences are likely not due to the expression of the PAT protein and do not represent meaningful differences with respect to the potential for adverse impact to the environment (CFIA 1995a, 1995b, 1996a, 1996b, 1996c, 1996d, 1996e, 1996f, 1998a, 1998b, 1998c, 1999, 2002a, 2002b, 2005, 2006a, EC 1996, 1997, 1998, 2001, Japan BCH 1996a, 1996b, 1996c, 1997a, 1997b, 1997c, 1997d, 1997e, 1999a, 1999b, 1999c, 1999d, 2002, 2005, 2006a, 2006b, 2006c, 2006d, 2006e, 2006f, 2007a, 2007b, 2008, 2009, 2010, OGTR 2002, 2003, 2006, Philippines 2005, USDA APHIS 1995a, 1995c, 1995f, 1996b, 1996b, 1996c, 1996e, 1997c, 1997f, 1998a, 1998c, 1998e, 1998g, 1998i, 1998j, 1999a, 1999b, 1999c,
2001b, 2001c, 2002b, 2003c, 2004a, 2004c, 2005, 2006b, USEPA 2001, 2005, 2009a, 2009b.)
WeedIness In agrIcultural envIronments
All of the plant species that have been engineered to express PAT have some potential to “volunteer” as weeds in subsequent grow-ing seasons and demonstrate varying degrees of ability to persist in an agricultural environment (OECD 1997, 2000, 2001, 2003a, 2008, OGTR 2008). The characteristics that influence the ability of a plant to volunteer are largely the same as those for weediness in general, such as seed dormancy, shattering, and competitiveness (Baker 1974). The data available indicate there is no linkage be-tween PAT protein expression and any increased survival or over-wintering capacity that would alter the prevalence of volunteer plants in the subsequent growing season (CFIA 1995a, 1995b, 1996a, 1996b, 1996c, 1996d, 1996e, 1996f, 1998a, 1998b, 1998c, 1999, 2002a, 2002b, 2005, 2006, EC 1996, 1997, 1998, 2001, Japan BCH 1996a, 1996b, 1996c, 1997a, 1997b, 1997c, 1997d, 1997e, 1998, 1999a, 1999b, 1999c, 1999d, 2002, 2005, 2006a, 2006b, 2006c, 2006d, 2006e, 2007a, 2007b, 2008, 2009, 2010, OGTR 2002, 2003, 2006, Philippines 2005, USDA APHIS 1995a, 1995c, 1995f, 1996b, 1996b, 1996c, 1996e, 1997c, 1997f, 1998a, 1998c, 1998e, 1998g, 1998i, 1998j, 1999a, 1999b, 1999c, 2001b, 2001c, 2002b, 2003c, 2004a, 2004c, 2005, 2006b, USEPA 2001, 2005, 2009a, 2009b). Following-season volunteers expressing PAT may complicate volunteer management programs, particularly if different crop species expressing glufosinate tolerance are planted in consecutive rotations. Alternative options are available for man-aging glufosinate-tolerant volunteers, including the use of other herbicides and mechanical weed control (Beckie et al. 2004, OECD 1997, OECD 2000, OECD 2001, OECD 2003a, OECD 2008, OGTR 2008).
WeedIness In non-agrIcultural envIronments
The primary mechanisms by which PAT may be introduced into a non-agricultural environment are: (1) seed or propagule move-ment (which may include incidental release during transportation of commodities) and establishment of the GE plant outside of cul-tivated areas, and; (2) gene flow from the GE plant to a natural-ized (or feral) population of the same crop species or other sexually compatible relatives (Mallory-Smith and Zapiola, 2008). Risk as-sessments for GE plants expressing PAT have considered the po-tential impacts associated with both types of introduction (CFIA 1995a, 1995b, 1996a, 1996b, 1996c, 1996d, 1996e, 1996f, 1998a, 1998b, 1998c, 1999, 2002a, 2002b, 2005, 2006, EC 1996, 1997, 1998, 2001, Japan BCH 1996a, 1996b, 1996c, 1997a, 1997b, 1997c, 1997d, 1997e, 1998, 1999a, 1999b, 1999c, 1999d, 2002, 2005, 2006a, 2006b, 2006c, 2006d, 2006e, 2007a, 2007b, 2008, 2009, 2010, OGTR 2002, 2003, 2006, Philippines 2005, USDA APHIS 1995a, 1995c, 1995f, 1996b, 1996b, 1996c, 1996e, 1997c, 1997f, 1998a, 1998c, 1998e, 1998g, 1998i, 1998j, 1999a, 1999b, 1999c, 2001b, 2001c, 2002b, 2003c, 2004a, 2004c, 2005, 2006b, USEPA 2001, 2005, 2009a, 2009b).
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While all plants can be considered weeds in certain contexts, none of the crops for which glufosinate-tolerant GE lines are available are considered to be invasive or problematic weeds outside of agri-cultural systems. Most can persist under favorable conditions and they may at times require management, particularly when they vol-unteer in subsequent crops (OECD 1997, OECD 2000, OECD 2001, OECD 2003a, OECD 2008, OGTR 2008, USDA APHIS 2004d). Based on agronomic and compositional data showing that PAT does not have a significant impact on agronomic or compo-sitional traits (including those that are related to weediness), the evidence to date shows that expression of the PAT protein has not resulted in any altered potential for weediness for those GE plant events subjected to environmental risk assessment. PAT expression only affects the ability of the plant to survive if treated with glu-fosinate. Just as in agricultural environments, other management options to control glufosinate-tolerant plants in non-agricultural environments are available (Beckie et al. 2004, OECD 1997, OECD 2000, OECD 2001, OECD 2003a, OECD 2008, OGTR 2008).
movement oF the transgene to WIld relatIves
The movement of transgenes to wild relatives is pollen-mediated and the production of reproductively viable hybrids depends on the physical proximity and flowering synchrony of the GE plants to sexually compatible species. The evidence shows that expression of the PAT protein in a range of plant species has not resulted in any alteration to anticipated gene flow. However introgression of glu-fosinate tolerance into sexually compatible, weedy populations in agricultural or peri-agricultural ecosystems is possible and has the potential to raise management issues (Mallory-Smith and Zapiola 2008, Warwick et al. 2007). In at least one instance, a regulato-ry decision has geographically limited the release of a herbicide-tolerant GE plant: the environmental approval of B. rapa event ZSR500/502 (glyphosate resistance) was limited to the western region of Canada due to the presence of feral populations of B. rapa in eastern Canada where it is considered a weed of agriculture (CFIA 1998d). However, no such decisions have been made for plants expressing PAT that are glufosinate-resistant, and all of the publicly available regulatory decisions conclude that the movement of the pat gene to wild relatives is not a substantial risk for any of the GE plants that have been considered (CFIA 1995a, 1995b, 1996a, 1996b, 1996c, 1996d, 1996e, 1996f, 1998a, 1998b, 1998c, 1999, 2002a, 2002b, 2005, 2006a, EC 1996, 1997, 1998, 2001, Japan BCH 1996a, 1996b, 1996c, 1997a, 1997b, 1997c, 1997d, 1997e, 1999a, 1999b, 1999c, 1999d, 2002, 2005, 2006a, 2006b, 2006c, 2006d, 2006e, 2006f, 2007a, 2007b, 2008, 2009, 2010, OGTR 2002, 2003, 2006, Philippines 2005, USDA APHIS 1995a, 1995c, 1995f, 1996b, 1996b, 1996c, 1996e, 1997c, 1997f, 1998a, 1998c, 1998e, 1998g, 1998i, 1998j, 1999a, 1999b, 1999c, 2001b, 2001c, 2002b, 2003c, 2004a, 2004c, 2005, 2006b, USEPA 2001, 2005, 2009a, 2009b).
adverse ImPacts on other organIsms In the receIvIng envIronment
The potential for PAT protein expression in GE plants to have an adverse impact on organisms has been considered in regula-tory risk assessments using a weight-of-evidence approach (CFIA 1995a, 1995b, 1996a, 1996b, 1996c, 1996d, 1996e, 1996f, 1998a, 1998b, 1998c, 1999, 2002a, 2002b, 2005, 2006a, EC 1996, 1997, 1998, 2001, Japan BCH 1996a, 1996b, 1996c, 1997a, 1997b, 1997c, 1997d, 1997e, 1999a, 1999b, 1999c, 1999d, 2002, 2005, 2006a, 2006b, 2006c, 2006d, 2006e, 2006f, 2007a, 2007b, 2008, 2009, 2010, OGTR 2002, 2003, 2006, Philippines 2005, USDA APHIS 1995a, 1995c, 1995f, 1996b, 1996b, 1996c, 1996e, 1997c, 1997f, 1998a, 1998c, 1998e, 1998g, 1998i, 1998j, 1999a, 1999b, 1999c, 2001b, 2001c, 2002b, 2003c, 2004a, 2004c, 2005, 2006b, USEPA 2001, 2005, 2009a, 2009b). These risk assessments have generally considered the potential for the novel protein to be toxic to other organisms, as well as the history of prior environmental exposure to the protein. Toxic proteins are known to act acutely (Sjoblad et al. 1992). Acute, intravenous toxicity experiments in mice show the PAT protein has no toxicity even at doses much higher than would be encountered due to environmental exposure to GE plants expressing the PAT protein (Herouet et al. 2005). In addition, the PAT protein shows no homology to known toxins or allergens and is rapidly digested in experiments simulating gastric environment (Herouet et al. 2005). The Streptomyces bacteria which are the source of PAT proteins are widespread in environments around the world, and additional species of Streptomyces are known to possess similar enzymatic activity, indicating that PAT protein homologs are likely ubiquitous in the environment and regula-tory decisions have concluded that exposure to PAT proteins from GE plants does not represent a potential for adverse impacts on other organisms (Herouet et al. 2005, CFIA 1995a, 1995b, 1996a, 1996b, 1996c, 1996d, 1996e, 1996f, 1998a, 1998b, 1998c, 1999, 2002a, 2002b, 2005, 2006a, EC 1996, 1997, 1998, 2001, Japan BCH 1996a, 1996b, 1996c, 1997a, 1997b, 1997c, 1997d, 1997e, 1999a, 1999b, 1999c, 1999d, 2002, 2005, 2006a, 2006b, 2006c, 2006d, 2006e, 2006f, 2007a, 2007b, 2008, 2009, 2010, OGTR 2002, 2003, 2006, Philippines 2005, USDA APHIS 1995a, 1995c, 1995f, 1996b, 1996b, 1996c, 1996e, 1997c, 1997f, 1998a, 1998c, 1998e, 1998g, 1998i, 1998j, 1999a, 1999b, 1999c, 2001b, 2001c, 2002b, 2003c, 2004a, 2004c, 2005, 2006b, USEPA 2001, 2005, 2009a, 2009b).
Risk assessors have also considered whether the introduction of PAT proteins into GE plants might lead to changes in the plant that would adversely impact other organisms. Phenotypic charac-terization (see above) as well as compositional analyses (see below) and nutritional analyses show that the introduction of PAT pro-teins has not had any unanticipated effects on characteristics of GE plants that might impact other organisms (ANZFA 2000, 2001a, 2001b, 2002, CFIA 1995a, 1995b, 1996a, 1996b, 1996c, 1996d, 1996e, 1996f, 1998a, 1998b, 1998c, 1999, 2002a, 2002b, 2004, 2005, 2006a, 2006b, EC 1996, 1997, 1998, 2001, EFSA 2005, 2006, 2008a, 2008b, 2009a, 2009b, FSANZ 2003, 2004a, 2004b, 2005a, 2005b, 2008, Japan BCH 1996a, 1996b, 1996c, Health Canada 2006a, 2006b, Japan BCH 1997a, 1997b, 1997c, 1997d, 1997e, 1999a, 1999b, 1999c, 1999d, 2002, 2003, 2005, 2006a,
7
2006b, 2006c, 2006d, 2006e, 2006f, 2007a, 2007b, 2008, 2009, 2010, OGTR 2003, 2006, Philippines 2005, USDA APHIS 1994a, b, 1995b, 1995d, 1995e, 1996a, 1996d, 1997a, 1997b, 1997d, 1997e, 1998b, 1998d, 1998f, 1998h, 1998k, 1998l, 2000, 2001a, 2002a, 2002c, 2003a, 2003b, 2004b, 2004d, 2006a, USEPA 2001, 2005, 2009a, 2009b). Based on experimental evidence that PAT proteins are not toxic and the observation that exposure to PAT is widespread in the environment, regulatory authorities have con-cluded that the expression of PAT in GE plants does not have any meaningful potential to adversely impact other organisms (CFIA 1995a, 1995b, 1996a, 1996b, 1996c, 1996d, 1996e, 1996f, 1998a, 1998b, 1998c, 1999, 2002a, 2002b, 2005, 2006a, EC 1996, 1997, 1998, 2001, Japan BCH 1996a, 1996b, 1996c, 1997a, 1997b, 1997c, 1997d, 1997e, 1999a, 1999b, 1999c, 1999d, 2002, 2005, 2006a, 2006b, 2006c, 2006d, 2006e, 2006f, 2007a, 2007b, 2008, 2009, 2010, OGTR 2002, 2003, 2006, Philippines 2005, USDA APHIS 1995a, 1995c, 1995f, 1996b, 1996b, 1996c, 1996e, 1997c, 1997f, 1998a, 1998c, 1998e, 1998g, 1998i, 1998j, 1999a, 1999b, 1999c, 2001b, 2001c, 2002b, 2003c, 2004a, 2004c, 2005, 2006b, USEPA 2001, 2005, 2009a, 2009b).
comPosItIonal analysIs oF Pat-exPressIng Plants
Detailed compositional analysis is a scientifically rigorous com-ponent of the characterization of GE plants and is a regulatory requirement for GE food and feed safety approvals (OECD 1992; WHO 1995, FAO/WHO 1996, EFSA 2006A, Codex 2003a, 2003b). The choice of analyses conducted depends on the nature of the product and its intended uses. Although compositional anal-ysis is not typically required for environmental risk assessments, it is often considered in the context of demonstrating whether or not there have been unanticipated changes to the GE plant (CFIA 1995a, 1995b, 1996a, 1996b, 1996c, 1996d, 1996e, 1996f, 1998a, 1998b, 1998c, 1999, 2002a, 2002b, 2005, 2006a, EC 1996, 1997, 1998, 2001, Japan BCH 1996a, 1996b, 1996c, 1997a, 1997b, 1997c, 1997d, 1997e, 1999a, 1999b, 1999c, 1999d, 2002, 2005, 2006a, 2006b, 2006c, 2006d, 2006e, 2006f, 2007a, 2007b, 2008, 2009, 2010, OGTR 2002, 2003, 2006, Philippines 2005, USDA APHIS 1995a, 1995c, 1995f, 1996b, 1996b, 1996c, 1996e, 1997c, 1997f, 1998a, 1998c, 1998e, 1998g, 1998i, 1998j, 1999a, 1999b, 1999c, 2001b, 2001c, 2002b, 2003c, 2004a, 2004c, 2005, 2006b, USEPA 2001, 2005, 2009a, 2009b). GE plants expressing PAT have undergone a variety of compositional analyses, including for proximate (protein, fat, amino acid, fiber, ash) as well as for nu-tritional components and known toxicants or antinutrients (such as gossypol in cotton or glucosinolates in canola) (ANZFA 2000, 2001a, 2001b, 2002, CFIA 1995a, 1995b, 1996a, 1996b, 1996c, 1996d, 1996e, 1996f, 1998a, 1998b, 1998c, 1999, 2002a, 2002b, 2004, 2005, 2006a, 2006b, EC 1996, 1997, 1998, 2001, EFSA 2005, 2006, 2008a, 2008b, 2009a, 2009b, FSANZ 2003, 2004a, 2004b, 2005a, 2005b, 2008, OGTR 2003, 2006, Philippines 2005, USDA APHIS 1994a, 1994b, 1995b, 1995d, 1995e, 1996a, 1996d, 1997a, 1997b, 1997d, 1997e, 1998b, 1998d, 1998f, 1998h, 1998k, 1998l, 2000, 2001a, 2002a, 2002c, 2003a, 2003b, 2004b, 2004d, 2006a, USEPA 2001, 2005, 2009a, 2009b). Although statistically significant differences between the composition of GE plants and
their non-transformed counterparts have been reported, these dif-ferences have not been attributed to expression of the PAT protein, and subsequent regulatory decisions have concluded that the com-position of GE plants expressing PAT is not meaningfully different with respect to potential impact on the environment (CFIA 1995a, 1995b, 1996a, 1996b, 1996c, 1996d, 1996e, 1996f, 1998a, 1998b, 1998c, 1999, 2002a, 2002b, 2005, 2006a, EC 1996, 1997, 1998, 2001, Japan BCH 1996a, 1996b, 1996c, 1997a, 1997b, 1997c, 1997d, 1997e, 1999a, 1999b, 1999c, 1999d, 2002, 2005, 2006a, 2006b, 2006c, 2006d, 2006e, 2006f, 2007a, 2007b, 2008, 2009, 2010, OGTR 2002, 2003, 2006, Philippines 2005, USDA APHIS 1995a, 1995c, 1995f, 1996b, 1996b, 1996c, 1996e, 1997c, 1997f, 1998a, 1998c, 1998e, 1998g, 1998i, 1998j, 1999a, 1999b, 1999c, 2001b, 2001c, 2002b, 2003c, 2004a, 2004c, 2005, 2006b, USEPA 2001, 2005, 2009a, 2009b).
conclusIon
The PAT protein expressed in GE plants is encoded by one of the homologous genes pat or bar, isolated from the related bacteria Streptomyces viridochromogenes or Streptomyces hygroscopicus, respec-tively. Environmental release approvals have been granted for 8 spe-cies of plants expressing PAT proteins in 11 different countries in-cluding at least 38 separate transformation events. Data from regu-latory submissions and peer-reviewed literature show that the PAT protein expressed in GE plants has negligible impact on the pheno-type of those plants, beyond conferring tolerance to the herbicide glufosinate. Risk assessments associated with regulatory review of these plants for use in the environment show that expression of PAT does not alter the potential for persistence or spread of GE plants in the environment, does not alter the reproductive biology or potential for gene flow, and does not increase the risks for ad-verse effects to other organisms. Although the introduction of PAT to GE plants has the potential to complicate the management of herbicide-tolerant volunteers or weedy relatives in agriculture, the evidence does not indicate that expression of PAT has impacted the effectiveness or availability of alternative control measures such as other herbicides or mechanical weed control. Taken together, these regulatory analyses support the conclusion that, for the species and environments that have been considered to date, the expression of the PAT protein in GE plants does not present any meaningful risk to the environment.
reFerences
Journal Articles and Books
Baker H.G. (1974). The evolution of weeds. Annual Review of Ecology and Systematics 5:1-24.
Beckie H.J., Seguin-Swartz G., Nair H., Warwick S.I., and Johnson E. (2004). Multiple herbicide-resistant canola can be controlled by alternative herbicides. Weed Science 52 (1):152-157.
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De Block M., Botterman J., Vandewiele M., Dockx J., Thoen C., Gossele V., Movva N.R., Thompson C., van Montagu M., and Leemans J. (1987). Engineer-ing herbicide resistance in plants by expression of a detoxifying enzyme. The EMBO Journal 6(9):2513-2518.
Droge-Laser W., Siemeling U.,Puhler A., and Broer I. (1994). The metabolites of the herbicide L-Phosphinothricin (Glufosinate): Identification, stability and mo-bility in transgenic, herbicide-resistant and untransformed plants. Plant Physiol-ogy 105:159-166.
Herouet C., Esdaile D. J., Mallyon B. A., Debruyne E., Schulz A., Currier T., Hendrickx K., van der Klis R-J., and Rouan D. (2005). Safety evaluation of the phosphinothricin acetyltransferase proteins encoded by the pat and bar se-quences that confer tolerance to glufosinate-ammonium herbicide in transgenic plants. Regulatory Toxicology and Pharmacology 41:134-149.
Hoerlein G. (1994). Glufosinate (Phosphinothricin), a Natural Amino Acid with Unexpected Herbicidal Properties. Reviews of Environmental Contamination and Toxicology 138:73-145.
Mallory-Smith C. and Zapiola M. (2008). Gene flow from glyphosate-resistant crops. Pest Management Science 64:428-440.
Schwarz D., Berger S., Heinzelmann E., Muschko K., Welzel K., and Wohlleben W. (2004). Biosynthetic gene cluster of the herbicide phosphinothricin tripep-tide from Streptomyces viridochromogenes Tu494. Applied and Environmental Microbiology 70(12):7093-7102.
Sjoblad R.D., McClintock J.T. and Engler R. (1992). Toxicological considera-tions for protein components of biological pesticide products. Regulatory Toxi-cology and Pharmacology 15: 3-9.
Thompson C. J., Movva N. R., Tizard R., Crameri R, Davies J. E., Lauwereys M., and Botterman J. (1987). Characterization of the herbicide-resistance gene bar from Streptomyces hygroscopicus. The EMBO Journal 6(9):2519-2523.
Warwick, S.I., Legere, A., Simard, M.-J., and James, T. (2008). Do escaped transgenes persist in nature? The case of an herbicide resistance transgene in a weedy Brassica rapa population. Molecular Ecology. 17:1387-1395.
Wehrmann A., van Vliet A., Opsomer C., Botterman J., and Schulz A. (1996). The similarities of bar and pat gene products make them equally applicable for plant engineers. Nature Biotechnology 14:1274-1278.
Wohlleben W., Arnold W., Broer I. Hillermann D., Strauch E., and Puhler A. (1988). Nucleotide sequence of the phosphinothricin N-acetyltransferase gene from Streptomyces viridochromogenes Tu494 and its expression in Nicotiana tabacum. Gene 70:25-37.
Regulatory Citations
ANZFA (2000). Draft Risk Analysis Report: Food derived from insect-protected Bt-176 corn. Australia New Zealand Food Authority, Canberra, Australia. http://www.foodstandards.gov.au/_srcfiles/A385%20FA.pdf
ANZFA (2001a). Draft Risk Assessment Report: Food derived from insect-protected, herbicide-tolerant BT11 corn. Australia New Zealand Food Authority, Canberra, Australia. http://www.foodstandards.gov.au/_srcfiles/A386_IR.pdf
ANZFA (2001b). Risk assessment for application A375: food derived from glu-fosinate tolerant maize line T25. Australia New Zealand Food Authority, Can-berra, Australia. http://www.foodstandards.gov.au/_srcfiles/A375%20Final%20AR.pdf
ANZFA (2002). Final assessment report (inquiry - section 17) application A380 food from insect-protected and glufosinate ammonium-tolerant DBTt418 corn. Australia New Zealand Food Authority, Canberra, Australia. http://www.food-standards.gov.au/_srcfiles/A380_Final_Assessment_Report.pdf
CFIA (1995a). Decision document DD95-01: Determination of environmental safety of Agrevo Canada Inc.’s glufosinate ammonium-tolerant canola. Cana-dian Food Inspection Agency (CFIA) Ottawa, Canada. http://www.inspection.gc.ca/english/plaveg/bio/dd/dd9501e.shtml
CFIA (1995b). Decision document DD95-04: Determination of environmental safety of Plant Genetic systems Inc. (PGS) novel hybridization system for can-ola (Brassica napus L.) (MS1, RF1). Canadian Food Inspection Agency (CFIA) Ottawa, Canada. http://www.inspection.gc.ca/english/plaveg/bio/dd/dd9504e.shtml
CFIA (1996a). Decision document DD96-09: Determination of environmental safety of event 176 Bt corn (Zea mays L.) developed by Ciba Seeds and Myco-gen Corporation. Canadian Food Inspection Agency (CFIA) Ottawa, Canada. http://www.inspection.gc.ca/english/plaveg/bio/dd/dd9609e.shtml
CFIA (1996b). Decision document DD96-11: Determination of environmen-tal safety of Agrevo Canada Inc.’s glufosinate ammonium-tolerant canola line HCN28(T45). Canadian Food Inspection Agency (CFIA) Ottawa, Canada. http://www.inspection.gc.ca/english/plaveg/bio/dd/dd9611e.shtml
CFIA (1996c). Decision Document DD96-12: Determination of environmental safety of Northrup King Seeds’ european corn borer (ECB) resistant corn (Zea mays L.). Canadian Food Inspection Agency (CFIA) Ottawa, Canada. http://www.inspection.gc.ca/english/plaveg/bio/dd/dd9612e.shtml
CFIA (1996d). Decision document 96-15: CFIA (1996c). Determination of envi-ronmental safety of Dekalb Canada Inc.’s glufosinate ammonium-tolerant corn line DLL25. Canadian Food Inspection Agency (CFIA) Ottawa, Canada. http://www.inspection.gc.ca/english/plaveg/bio/dd/dd9615e.shtml
CFIA (1996e). Decision document 96-16: Determination of environmental safe-ty of Plant Genetic Systems Inc.’s (PGS) male sterile corn (Zea mays L.) line MS3. Canadian Food Inspection Agency (CFIA) Ottawa, Canada. http://www.inspection.gc.ca/english/plaveg/bio/dd/dd9616e.shtml
CFIA (1996f). Decision document 96-17: Determination of environmental safety of Plant Genetic Systems Inc.’s (PGS) novel hybridization system for rapeseed (Brassica napus L.) (MS8xRF3). Canadian Food Inspection Agency (CFIA) Ottawa, Canada. http://www.inspection.gc.ca/english/plaveg/bio/dd/dd9617e.shtml
CFIA (1998a). Decision Document 98-22: Determination of the safety of AgrEvo Canada Inc.’s glufosinate ammonium tolerant corn (Zea mays) lines, T14 and T25. Canadian Food Inspection Agency (CFIA) Ottawa, Canada. http://www.inspection.gc.ca/english/plaveg/bio/dd/dd9822e.shtml
CFIA (1998b). Decision Document DD98-23: Decision document 98-23: deter-mination of environmental safety of Dekalb Genetics Corporation’s European corn borer (ECB) resistant corn (Zea mays L.) line DBT418. Canadian Food In-spection Agency (CFIA) Ottawa, Canada. http://www.inspection.gc.ca/english/plaveg/bio/dd/dd9823e.shtml
CFIA (1998c). Decision Document DD98-28: Determination of the safety of AgrEvo Canada Inc.’s glufosinate ammonium herbicide-tolerant Brassica rapa canola line HCR-1. Canadian Food Inspection Agency (CFIA) Ottawa, Canada. http://www.inspection.gc.ca/english/plaveg/bio/dd/dd9828e.shtml
CFIA (1998d). Determination of the safety of Monsanto Canada Inc.’s roundup herbicide-tolerant Brassica rapa canola lines ZSR500, ZSR502, and ZSR503. Canadian Food Inspection Agency (CFIA), Ottawa, Canada. http://www.inspec-tion.gc.ca/english/plaveg/bio/dd/dd9821e.shtml
CFIA (1999). Decision Document DD99-30: Determination of the environ-mental safety of AgrEvo Canada Inc.’s glufosinate ammonium tolerant soybean (Glycine max) A2704-12. Canadian Food Inspection Agency (CFIA) Ottawa, Canada. http://www.inspection.gc.ca/english/plaveg/bio/dd/dd9930e.shtml
CFIA (2002a). Decision document DD2002-39: Determination of the safety of Aventis CropsCience Canada Inc’s glufosinate ammonium-tolerant sugar beet (Beta vulgaris) lines derived from event T120-7. Canadian Food Inspection Agency (CFIA) Ottawa, Canada. http://www.inspection.gc.ca/english/plaveg/bio/dd/dd0239e.shtml
CFIA (2002b) Decision Document DD2002-41: Determination of the Safety of Dow AgroSciences Canada Inc. and Pioneer Hi-Bred International’s Insect Re-sistant and Glufosinate - Ammonium Tolerant Corn (Zea mays L.) Line 1507. Canadian Food Inspection Agency (CFIA) Ottawa, Canada. http://www.inspec-tion.gc.ca/english/plaveg/bio/dd/dd0241e.shtml
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CFIA (2004). Decision Document DD2004-49: Determination of the Safety of Bayer CropScience’s Herbicide Tolerant LibertyLink® Cotton Event LLcot-ton25 (Gossypium hirsutum L.). Canadian Food Inspection Agency (CFIA) Otta-wa, Canada. http://www.inspection.gc.ca/english/plaveg/bio/dd/dd0449e.shtml
CFIA (2005). Decision Document DD2005-55: Determination of the safety of Dow AgroSciences Canada Inc. and Pioneer Hi-Bred Production Inc.’s insect resistant and glufosinate-ammonium herbicide tolerant corn (Zea mays L.) Line 59122. Canadian Food Inspection Agency (CFIA) Ottawa, Canada. http://www.inspection.gc.ca/english/plaveg/bio/dd/dd0555e.shtml
CFIA (2006a). Decision document DD2006-59: Determination of the safety of Dow AgroSciences Canada Inc.’s insect resistant and glufosinate - ammonium tolerant corn (Zea mays L.) Event DAS-06275-8. Canadian Food Inspection Agency (CFIA) Ottawa, Canada. http://www.inspection.gc.ca/english/plaveg/bio/dd/dd0659e.shtml
CFIA (2006b). Decision Document DD2006-58: Determination of the safety of Bayer CropScience’s glufosinate ammonium tolerant rice (Oryza sativa) Event LLrice62. Canadian Food Inspection Agency (CFIA) Ottawa, Canada. http://www.inspection.gc.ca/english/plaveg/bio/dd/dd0658e.shtml
European Commission (1996). 96/424/EC: Commission Decision of 20 May 1996 concerning the placing on the market of genetically modified male sterile chicory (Cichorium intybus L.) with partial tolerance to the herbicide glufosinate ammonium pursuant to Council Directive 90/220/EEC. Official journal of the European Union NO. L 175 , 13/07/1996 P. 0025 – 0026. http://www.biosafety.be/GB/Dir.Eur.GB/Market/96_424/96_424.html
European Commission (1997). 97/98/EC: Commission Decision of 23 January 1997 concerning the placing on the market of genetically modified maize (Zea mays L.) with the combined modification for insecticidal properties conferred by the Bt-endotoxin gene and increased tolerance to the herbicide glufosinate ammonium pursuant to Council Directive 90/220/EEC. Official journal of the European Union NO. L 031 , 01/02/1997 P. 0069 – 0070. http://www.biosafety.be/PDF/97_98.pdf
European Commission (1998). 98/293/EC: Commission Decision of 22 April 1998 concerning the placing on the market of genetically modified maize (Zea mays L. T25), pursuant to Council Directive 90/220/EEC (Text with EEA rele-vance). Official Journal L 131 , 05/05/1998 p. 0030 – 0031. http://eur-lex.europa.eu/LexUriServ/LexUriServ.do?uri=OJ:L:1998:131:0030:0031:EN:PDF
European Commission (2001). Opinion of the scientific committee on plants re-garding “submission for placing on the market of glufosinate tolerant maize (Zea mays) transformation eventT25” by the AgrEvo company now Aventis Crop-science (Notification C/F/95/12/07). European Commission, Brussels, Belgium. http://ec.europa.eu/food/fs/sc/scp/out04_en.html
EFSA (2005) For citation purposes: Opinion of the Scientific Panel on Genetical-ly Modified Organisms on an application (reference EFSA-GMONL-2004-02) for the placing on the market of insect-tolerant genetically modified maize 1507, for food use, under Regulation (EC) No 1829/2003 from Pioneer Hi-Bred In-ternational/Mycogen Seeds, The EFSA Journal (2005) 182, 1-22. http://www.europabio.org/InfoOperators/EFSA%20Opinion1507_190105.pdf
EFSA (2006). Opinion of the Scientific Panel on Genetically Modified Organ-isms on an application (Reference EFSA-GMO-NL-2005-13) for the placing on the market of glufosinate-tolerant genetically modified LLCotton25, for food and feed uses, and import and processing under Regulation (EC) No 1829/2003 from Bayer CropScience, The EFSA Journal (2006) 429, 1-19. http://www.efsa.europa.eu/en/scdocs/doc/429.pdf
EFSA (2008a). Opinion of the Scientific Panel on Genetically Modified Organ-isms on an application (Reference EFSA-GMO-UK-2005-25) for the placing on the market of glufosinate-tolerant oilseed rape T45 for food and feed uses, import and processing and renewal of the authorization of oilseed rape T45 as existing products, both under Regulation (EC) 1829/2003 from Bayer Crop-Science, The EFSA Journal (2008) 635, 1-22. http://www.efsa.europa.eu/en/scdocs/doc/635.pdf
EFSA (2008b)Scientific Opinion of the Panel on Genetically Modified Organ-isms on a request from Pioneer Hi-Bred International on the authorisation for the placing on the market of the insect-resistant and herbicide-tolerant genetically modified maize 59122 x NK603, for food and feed uses, and import and process-ing under Regulation (EC) No 1829/2003. The EFSA Journal (2008) 874, 1-34. http://www.efsa.europa.eu/en/scdocs/doc/874.pdf
EFSA (2009a). Scientific Opinion of the Panel on Genetically Modified Organ-isms on an application (Reference EFSA-GMO-UK-2005-21) for the placing on the market of insect-resistant and herbicide-tolerant genetically modified maize 59122 x 1507 x NK603 for food and feed uses, and import and processing under Regulation (EC) No 1829/2003 from Pioneer Hi-Bred International, Inc. The EFSA Journal (2009) 1050, 1-32. http://cera-gmc.org/docs/decdocs/09-235-005.pdf
EFSA (2009b). Opinion of the Scientific Panel on Genetically Modified Organ-isms on an application (Reference EFSA-GMO-NL-2005-15) for the placing on the market of the insect-resistant and herbicide-tolerant genetically modi-fied maize 1507 x 59122, for food and feed uses, import and processing under Regulation (EC) No 1829/2003 from Mycogen Seeds, c/o Dow AgroSciences LLC and Pioneer Hi-Bred International, Inc. as represented by Pioneer Over-seas Corporation, The EFSA Journal 1074, 1-28. http://www.efsa.europa.eu/en/scdocs/scdoc/1074.htm
FSANZ (2003). Final assessment report, application A446: food derived from insect-protected and glufosinate ammonium-tolerant corn line 1507. Food Standards Australia New Zealand (FSANZ), Canberra, Australia. http://www.foodstandards.gov.au/_srcfiles/ACF18.pdf
FSANZ (2004a). Final Assessment Report: Application A481- Food derived from glufosinate ammonium tolerant soybean lines A2704-12 and 15547-127. Food Standards Australia New Zealand (FSANZ), Canberra, Australia. http://www.foodstandards.govt.nz/_srcfiles/A481_GM_soy_FAR.pdf
FSANZ (2004b). Final Assessment Report: Application A518 - Food derived from insect-protected herbicide tolerant cotton line MXB-13. Food Standards Australia New Zealand (FSANZ), Canberra, Australia. http://www.foodstand-ards.govt.nz/_srcfiles/A518_GM_Cotton_FAR_FINAL.doc
FSANZ (2005a). Final Assessment Report: Application A533 - Food Derived from Glufosinate Ammonium-Tolerant Cotton Line LL25. Food Standards Aus-tralia New Zealand (FSANZ), Canberra, Australia. http://www.foodstandards.gov.au/_srcfiles/FAR_A533%20GM%20cotton.pdf
FSANZ (2005b). Final Assessment Report: Application A543 - Food Derived from Insect-Protected Glufosinate Ammonium-Tolerant Corn Line 59122-7. Food Standards Australia New Zealand (FSANZ), Canberra, Australia. http://www.foodstandards.gov.au/_srcfiles/A543_GM_Corn_DAR2.pdf
FSANZ (2008). Final Assessment Report: Application A589 - Food derived from glufosinate ammonium-tolerant rice LLRICE62. Food Standards Australia New Zealand (FSANZ), Canberra, Australia. http://www.foodstandards.gov.au/_srcfiles/A589_GM_LLRICE62_FAR.pdf
Health Canada (2006a). Novel Food Information; Glufosinate Tolerant Rice Event LLRICE62. Health Canada, Ottawa, Canada. http://www.hc-sc.gc.ca/fn-an/gmf-agm/appro/nf-an90decdoc-eng.php
Health Canada (2006b). Novel Food Information: (mo)Cry1F Insect Resistant, Glufosinate Tolerant Corn Event TC6275. Health Canada, Ottawa, Canada. http://www.hc-sc.gc.ca/fn-an/gmf-agm/appro/nf-an111decdoc-eng.php
Japanese Biosafety Clearing House (1996a). Outline of the biological diversity risk assessment report: Type 1 use approval for Topas 19/2 (HCN92). Japan Bi-osafety Clearing House (BCH). Tokyo, Japan.
Japanese Biosafety Clearing House (1996b). Outline of the biological diversity risk assessment report: Type 1 use approval for ACS-BN004-7×ACS-BN001-4 (Ms1, RF1). Japan Biosafety Clearing House (BCH). Tokyo, Japan.
Japanese Biosafety Clearing House (1996c). Outline of the biological diversity risk assessment report: Type 1 use approval for lepidopteran resistant, herbicide tolerant maize SYN-BT011-1. Japan Biosafety Clearing House (BCH). Tokyo, Japan.
10
Japanese Biosafety Clearing House (1997a). Outline of the biological diversity risk assessment report: Type 1 use approval for ACS-BN004-7×ACS-BN002-5 (MS1, RF2). Japan Biosafety Clearing House (BCH). Tokyo, Japan.
Japanese Biosafety Clearing House (1997b). Outline of the biological diversity risk assessment report: Type 1 use approval for ACS-BN008-2 (HCN28). Japan Biosafety Clearing House (BCH). Tokyo, Japan.
Japanese Biosafety Clearing House (1997c). Outline of the biological diversity risk assessment report: Type 1 use approval for T14. Japan Biosafety Clearing House (BCH). Tokyo, Japan.
Japanese Biosafety Clearing House (1997d). Outline of the biological diversity risk assessment report: Type 1 use approval for T25. Japan Biosafety Clearing House (BCH). Tokyo, Japan.
Japanese Biosafety Clearing House (1997e). Outline of the biological diversity risk assessment report: Type 1 use approval for SYN-EV176-9. Japan Biosafety Clearing House (BCH). Tokyo, Japan.
Japanese Biosafety Clearing House (1999a). Outline of the biological diversity risk assessment report: Type 1 use approval for ACS-BN005-8×ACS-BN003-6 (MS8x RF3). Japan Biosafety Clearing House (BCH). Tokyo, Japan.
Japanese Biosafety Clearing House (1999b). Outline of the biological diversity risk assessment report: Type 1 use approval for soybean A2704-12. Japan Bi-osafety Clearing House (BCH). Tokyo, Japan.
Japanese Biosafety Clearing House (1999c). Outline of the biological diversity risk assessment report: Type 1 use approval for DLL25. Japan Biosafety Clear-ing House (BCH). Tokyo, Japan.
Japanese Biosafety Clearing House (1999d). Outline of the biological diversity risk assessment report: Type 1 use approval for maize DBT418 (DKB-89614-9). Japan Biosafety Clearing House (BCH). Tokyo, Japan.
Japanese Biosafety Clearing House (2002). Outline of the biological diversity risk assessment report: Type 1 use approval for cotton DAS-01507-1. Japan Bi-osafety Clearing House (BCH). Tokyo, Japan.
Japanese Biosafety Clearing House (2003). Outline of the biological diversity risk assessment report: Type 1 use approval for T25 x MON810 maize. Japan Biosafety Clearing House (BCH). Tokyo, Japan.
Japanese Biosafety Clearing House (2005). Outline of the biological diversity risk assessment report: Type 1 use approval for DAS-01507-1 x MON-00603-6. Japan Biosafety Clearing House (BCH). Tokyo, Japan.
Japanese Biosafety Clearing House (2006a). Outline of the biological di-versity risk assessment report: Type 1 use approval for cotton LLCotton25 x MON15985 (ACS-GH001-3 x MON-15985-7). Japan Biosafety Clearing House (BCH). Tokyo, Japan.
Japanese Biosafety Clearing House (2006b). Outline of the biological diversity risk assessment report: Type 1 use approval for DAS-59122-7. Japan Biosafety Clearing House (BCH). Tokyo, Japan.
Japanese Biosafety Clearing House (2006c). Outline of the biological diversity risk assessment report: Type 1 use approval for DAS-59122-7 x NK603. Japan Biosafety Clearing House (BCH). Tokyo, Japan.
Japanese Biosafety Clearing House (2006d). Outline of the biological diver-sity risk assessment report: Type 1 use approval for DAS-59122-7 x TC1507 x NK603. Japan Biosafety Clearing House (BCH). Tokyo, Japan.
Japanese Biosafety Clearing House (2006e). Outline of the biological diversity risk assessment report: Type 1 use approval for TC1507 x DAS-59122-7. Japan Biosafety Clearing House (BCH). Tokyo, Japan.
Japanese Biosafety Clearing House (2006f). Outline of the biological diversity risk assessment report: Type 1 use approval for soybean A5547-127. Japan Bi-osafety Clearing House (BCH). Tokyo, Japan.
Japanese Biosafety Clearing House (2007a). Outline of the biological diversity risk assessment report: Type 1 use approval for soybean A5547-127. Japan Bi-osafety Clearing House (BCH). Tokyo, Japan.
Japanese Biosafety Clearing House (2007b). Outline of the biological diversity risk assessment report: Type 1 use approval for SYN-BT011-1 x MON-00021-9. Japan Biosafety Clearing House (BCH). Tokyo, Japan.
Japanese Biosafety Clearing House (2008). Outline of the biological diversity risk assessment report: Type 1 use approval for TC6275, (DAS-06275-8). Japan Biosafety Clearing House (BCH). Tokyo, Japan.
Japanese Biosafety Clearing House (2009). Outline of the biological diversity risk assessment report: Type 1 use approval for SmartStax maize. Japan Biosafe-ty Clearing House (BCH). Tokyo, Japan.
Japanese Biosafety Clearing House (2010). Outline of the biological diversity risk assessment report: Type 1 use approval for NK603 x T25 maize. Japan Bi-osafety Clearing House (BCH). Tokyo, Japan.
OECD (1992). Recombinant DNA safety considerations. Organization for Eco-nomic Cooperation and Development (OECD), Paris, France.
OECD (1993). Safety considerations for biotechnology: scale-up of crop plants. Organization for Economic Cooperation and Development (OECD), Paris, France.
OECD (1997). Consensus document on the biology of Brassica napus L. (oilseed rape). Organization for Economic Cooperation and Development (OECD), Paris,France.
OECD (1999a). Consensus document on general information concerning the genes and their enzymes that confer tolerance to phosphinothricin herbicide. Or-ganization for Economic Cooperation and Development (OECD), Paris, France. http://www.olis.oecd.org/olis/1999doc.nsf/LinkTo/env-jm-mono(99)13
OECD (1999b) Consensus Document on the Biology of Oryza sativa (Rice) No. 14, 1999 Organization for Economic Cooperation and Development (OECD), Paris, France. http://www.olis.oecd.org/olis/1999doc.nsf/LinkTo/env-jm-mo-no(99)26
OECD (2000) Consensus document on the biology of Glycine max (L.) Merr. Or-ganization for Economic Cooperation and Development (OECD), Paris, France.
OECD (2001). Consensus document on the biology of Beta vulgaris L. Organi-zation for Economic Cooperation and Development (OECD), Paris, France.
OECD (2002). Module II: Herbicide Biochemistry, Herbicide Metabolism and the Residues in Glufosinate-Ammonium (Phosphinothricin)-Tolerant Transgenic Plants. Organization for Economic Cooperation and Devel-opment (OECD), Paris, France. http://www.oecd.org/officialdocuments/displaydocumentpdf?cote=ENV/JM/MONO%282002%2914&doclanguage=en
OECD (2003a). Consensus document on the biology of Zea mays subsp. Mays. Organization for Economic Cooperation and Development (OECD), Paris, France.
OECD (2003b). Description of selected key generic terms used in chemical hazard/risk assessment. Organization for Economic Cooperation and Devel-opment (OECD), Paris. http://www.olis.oecd.org/olis/2003doc.nsf/LinkTo/NT00004772/$FILE/JT00152557.PDFOECD (2006). Points to consider for consensus documents on the biology of cultivated plants. Organization for Eco-nomic Cooperation and Development (OECD), Paris, France.
OECD (2008). Consensus document on the biology of cotton (Gossypium spp.). Organization for Economic Cooperation and Development (OECD), Paris, France.
OGTR (2003). Risk assessment and risk management plan: DIR 021/2002 Com-mercial release of genetically modified (InVigor hybrid) canola (MS1, RF1, RF2, RF3, T45, Topas 19/2, HCN92, MS1, MS8). Office of the Gene Technol-ogy Regulator (OGTR), Canberra, Australia. http://www.ogtr.gov.au/internet/ogtr/publishing.nsf/Content/dir021-2002
11
OGTR (2006). Risk assessment and risk management plan: DIR 062/2005 Com-mercial release of herbicide tolerant Liberty Link cotton for use in the Australian cropping system. Office of the Gene Technology Regulator (OGTR), Canberra, Australia. http://www.ogtr.gov.au/internet/ogtr/publishing.nsf/Content/dir062-2005
OGTR (2008). The biology of Gossypium hirsutum L. and Gossypium bar-badense L. Office of the gene technology regulatory (OGTR) Department of Health and Ageing, Canberra, Australia.
Philippines (2005). Determination of the Safety of Syngenta’s Corn BT11 (In-sect-resistant and herbicide-tolerant Corn) for Direct use as Food, Feed, and for Processing and for Propagation. Philippines Department of Agriculture, Bureau of Plant Industries, Manila, Philippines.
USDA APHIS (1994a). Petition for determination of nonregulated status of Ciba Seeds’ corn genetically engineered to express the Cry1A(b) protein from Bacil-lus thuringiensis subspecies kurstaki. United States Department of Agriculture, Animal and Plant Health Inspection Service, Washington D.C. http://www.aphis.usda.gov/brs/aphisdocs/94_31901p.pdf
USDA APHIS (1994b). Petition for determination of nonregulated status for glu-fosinate resistant corn transformation events T14 and T25. United States Depart-ment of Agriculture, Animal and Plant Health Inspection Service, Washington D.C. http://www.aphis.usda.gov/brs/aphisdocs/94_35701p.pdf
USDA APHIS (1995a). Environmental assessment and finding of no signifi-cant impact: Petition 94-319-01 for determination of nonregulated status for event 176 corn. United States Department of Agriculture, Animal and Plant Health Inspection Service, Washington D.C. http://www.aphis.usda.gov/brs/aphisdocs2/94_31901p_com.pdf
USDA APHIS (1995b). Petition for determination of nonregulated status: Glu-fosinate resistant corn line B16. United States Department of Agriculture, Ani-mal and Plant Health Inspection Service, Washington D.C. http://www.aphis.usda.gov/brs/aphisdocs/95_14501p.pdf
USDA APHIS (1995c). Environmental assessment and finding of no significant impact: Petition 95-145-01 for determination of nonregulated status for glufosi-nate resistant corn transformation line B16. United States Department of Agri-culture, Animal and Plant Health Inspection Service, Washington D.C. http://www.aphis.usda.gov/brs/aphisdocs2/95_14501p_com.pdf
USDA APHIS (1995d). Petition for determination of nonregulated status for: Insect protected corn (Zea mays L.) expressing the CryIA(b) gene from Bacil-lus thuringienisis var. kurstaki(Bt11). United States Department of Agriculture, Animal and Plant Health Inspection Service, Washington D.C. http://www.aphis.usda.gov/brs/aphisdocs/95_19501p.pdf
USDA APHIS (1995e). Petition for determination of nonregulated status for male sterile, glufosinate tolerant corn transformation event MS3. United States Department of Agriculture, Animal and Plant Health Inspection Service, Wash-ington D.C. http://www.aphis.usda.gov/brs/aphisdocs/95_22801p.pdf
USDA APHIS (1995f). Environmental assessment and finding of no significant impact: Petition 94-357-01 for determination of nonregulated status for glufosi-nate resistant corn transformation events T14 and T25. United States Department of Agriculture, Animal and Plant Health Inspection Service, Washington D.C. http://www.aphis.usda.gov/brs/aphisdocs2/94_35701p_com.pdf
USDA APHIS (1996a). Petition for determination of nonregulated status for glu-fosinate resistant soybean transformation events (W62, W98, A2704-12, A2704-21 and A5547-35). United States Department of Agriculture, Animal and Plant Health Inspection Service, Washington D.C. http://www.aphis.usda.gov/brs/aphisdocs/96_06801p.pdf
USDA APHIS (1996b). Environmental assessment and finding of no significant impact: AgroEvo USA company petition 96-068-01p for determination of non-regulated status for transgenic glufosinate resistant soybean (GRS) lines W62, W98, A2704-12, A2704-21 and A5547-35. United States Department of Agri-culture, Animal and Plant Health Inspection Service, Washington D.C. http://www.aphis.usda.gov/brs/aphisdocs2/96_06801p_com.pdf
USDA APHIS (1996c). Environmental assessment and finding of no sig-nificant impact: Petition 95-195-01 for determination of nonregulated status for Bt11 corn. United States Department of Agriculture, Animal and Plant Health Inspection Service, Washington D.C. http://www.aphis.usda.gov/brs/aphisdocs2/95_19501p_com.pdf
USDA APHIS (1996d). Petition for determination of nonregulated status: insect protected corn (Zea mays L.) with cryIA(c) gene from Bacillus thuringiensis subsp. kurstaki (DBT418). United States Department of Agriculture, Animal and Plant Health Inspection Service, Washington D.C. http://www.aphis.usda.gov/brs/aphisdocs/96_29101p.pdf
USDA APHIS (1996e). Environmental assessment and finding of no sig-nificant impact: Petition 95-228-01 for determination of nonregulated status for MS3 corn. United States Department of Agriculture, Animal and Plant Health Inspection Service, Washington D.C. http://www.aphis.usda.gov/brs/aphisdocs2/95_22801p_com.pdf
USDA APHIS (1997a). Petition for determination of nonregulated status: Glu-fosinate tolerant canola transformation event T45. United States Department of Agriculture, Animal and Plant Health Inspection Service, Washington D.C. http://www.aphis.usda.gov/brs/aphisdocs/97_20501p.pdf
USDA APHIS (1997b). Petition for determination of nonregulated status for Radicchio rosso lines with male sterility. United States Department of Agricul-ture, Animal and Plant Health Inspection Service, Washington D.C. http://www.aphis.usda.gov/brs/aphisdocs/97_14801p.pdf
USDA APHIS (1997c). Environmental assessment and finding of no significant impact: USDA/APHIS Petition 97-148-01p for determination of nonregulated status for Radiccio rosso lines designated as RM3-3, RM3-4, and RM3-6. United States Department of Agriculture, Animal and Plant Health Inspection Service, Washington D.C. http://www.aphis.usda.gov/brs/aphisdocs2/97_14801p_com.pdf
USDA APHIS (1997d). Petition for determination of nonregulated status for male sterile corn lines 676, 678, and 680 with resistance to glufosinate. United States Department of Agriculture, Animal and Plant Health Inspection Service, Washington D.C. http://www.aphis.usda.gov/brs/aphisdocs/97_34201p.pdf
USDA APHIS (1997e). Petition for determination of nonregulated status for Bt Cry9C insect resistant, glufosinate tolerant corn transformation event CBH-351. United States Department of Agriculture, Animal and Plant Health Inspection Service, Washington D.C. http://www.aphis.usda.gov/brs/aphisdocs/97_26501p.pdf
USDA APHIS (1997f). Environmental assessment and finding of no significant impact: USDA/APHIS Petition 96-291-01p for determination of nonregulated status for insect-protected corn line DBT418. United States Department of Ag-riculture, Animal and Plant Health Inspection Service, Washington D.C. http://www.aphis.usda.gov/brs/aphisdocs2/96_29101p_com.pdf
USDA APHIS (1998a). Environmental assessment and finding of no signifi-cant impact: AgrEvo USA Company Petition 97-205-01p for determination of nonregulated status for transgenic glufosinate tolerant canola transforma-tion (event T45). United States Department of Agriculture, Animal and Plant Health Inspection Service, Washington D.C. http://www.aphis.usda.gov/brs/aphisdocs2/97_20501p_com.pdf
USDA APHIS (1998b). Petition for determination of nonregulated status: Glu-fosinate tolerant sugar beet, transformation event T120-7. United States Depart-ment of Agriculture, Animal and Plant Health Inspection Service, Washington D.C. http://www.aphis.usda.gov/brs/aphisdocs/97_33601p.pdf
USDA APHIS (1998c). Environmental assessment and finding of no signifi-cant impact: AgrEvo USA Company Petition 97-336-01p for determination of nonregulated status for transgenic glufosinate tolerant sugar beet transforma-tion event T120-7. United States Department of Agriculture, Animal and Plant Health Inspection Service, Washington D.C. http://www.aphis.usda.gov/brs/aphisdocs2/97_33601p_com.pdf
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USDA APHIS (1998d). Application for an extension of determination of nonregulated status for glufosinate resistant soybean transformation events (A5547-127). United States Department of Agriculture, Animal and Plant Health Inspection Service, Washington D.C. http://www.aphis.usda.gov/brs/aphisdocs/98_01401p.pdf
USDA APHIS (1998e). Environmental assessment and finding of no significant impact: Request for extension of determination of nonregulated status for glu-fosinate resistant soybean transformation events (A5547-127). United States Department of Agriculture, Animal and Plant Health Inspection Service, Wash-ington D.C. http://www.aphis.usda.gov/brs/aphisdocs2/98_01401p_com.pdf
USDA APHIS (1998f). Application for an extension of determination of nonreg-ulated status for glufosinate resistant soybean transformation events (GU262). United States Department of Agriculture, Animal and Plant Health Inspection Service, Washington D.C. http://www.aphis.usda.gov/brs/aphisdocs/98_23801p.pdf
USDA APHIS (1998g). Environmental assessment and finding of no significant impact: application for an extension of determination of nonregulated status for glufosinate resistant soybean transformation events (GU262). United States De-partment of Agriculture, Animal and Plant Health Inspection Service, Washing-ton D.C. http://www.aphis.usda.gov/brs/aphisdocs2/98_23801p_com.pdf
USDA APHIS (1998h). Petition for determination of nonregulated status for LibertyLink rice transformation events LLRice06 and LLRice62. United States Department of Agriculture, Animal and Plant Health Inspection Service, Wash-ington D.C. http://www.aphis.usda.gov/brs/aphisdocs/98_32901p.pdf
USDA APHIS (1998i). Environmental assessment and finding of no significant impact: USDA/APHIS Petition 97-342-01p for determination of nonregulated status for genetically engineered corn lines 676, 678, and 680. United States Department of Agriculture, Animal and Plant Health Inspection Service, Wash-ington D.C. http://www.aphis.usda.gov/brs/aphisdocs2/97_34201p_com.pdf
USDA APHIS (1998j). Environmental assessment and finding of no sig-nificant impact: Petition 97-265-01p for determination of nonregulated sta-tus for Bt. Cry9C insect resistant, glufosinate tolerant corn transformation event CBH-351. United States Department of Agriculture, Animal and Plant Health Inspection Service, Washington D.C. http://www.aphis.usda.gov/brs/aphisdocs2/97_26501p_com.pdf
USDA APHIS (1998k). Request for extension of a determination of nonregu-lated status for a hybrid seed production system in corn based on male sterility and glufosinate tolerance as a marker (95-228-01p): event MS6. United States Department of Agriculture, Animal and Plant Health Inspection Service, Wash-ington D.C. http://www.aphis.usda.gov/brs/aphisdocs/98_34901p.pdf
USDA APHIS (1998l). Petition for determination of nonregulated status for In-Vigor hybrid canola transformation events MS8/RF3. United States Department of Agriculture, Animal and Plant Health Inspection Service, Washington D.C. http://www.aphis.usda.gov/brs/aphisdocs/98_27801p.pdf
USDA APHIS (1999a). Environmental assessment and finding of no significant impact: AgrEvo USA Petition 98-278-01p for determination of nonregulated sta-tus for canola transformation events MS8 and RF3 genetically engineered for pollination control and tolerance to glufosinate herbicide. United States Depart-ment of Agriculture, Animal and Plant Health Inspection Service, Washington D.C. http://www.aphis.usda.gov/brs/aphisdocs2/98_27801p_com.pdf
USDA APHIS (1999b). Environmental assessment and finding of no sig-nificant impact: AgrEvo USA Petition 98-329-01p for determination of non-regulated status for glufosinate tolerant rice transformation events LLRice06 and LLRice62. United States Department of Agriculture, Animal and Plant Health Inspection Service, Washington D.C. http://www.aphis.usda.gov/brs/aphisdocs2/98_32901p_com.pdf
USDA APHIS (1999c). Extension of determination of nonregulated status for corn genetically engineered for male sterility and glufosinate herbicide toler-ance as a marker (MS6). United States Department of Agriculture, Animal and Plant Health Inspection Service, Washington D.C. http://www.aphis.usda.gov/brs/aphisdocs2/98_34901p_com.pdf
USDA APHIS (2000). Petition for the determination of nonregulated status B.t. Cry1F insect resistant, glufosinate tolerant maize line. United States Department of Agriculture, Animal and Plant Health Inspection Service, Washington D.C. http://www.aphis.usda.gov/brs/aphisdocs/00_13601p.pdf
USDA APHIS (2001a). Application for an extension of the determination of nonregulated status for glufosinate-tolerant canola transformration (98-278-01p): MS1/RF1/RF2(01-206-01). United States Department of Agriculture, Ani-mal and Plant Health Inspection Service, Washington D.C. http://www.aphis.usda.gov/brs/aphisdocs/01_20601p.pdf
USDA APHIS (2001b). Extension of determination of nonregulated status for canola genetically engineered for male sterility, fertility restoration, and glufosi-nate herbicide tolerance (MS1, RF1, RF2). United States Department of Agricul-ture, Animal and Plant Health Inspection Service, Washington D.C. http://www.aphis.usda.gov/brs/aphisdocs2/01_20601p_com.pdf
USDA APHIS (2001c). Environmental assessment and finding of no signifi-cant impact: Approval of Mycogen Seeds c/o Dow AgroSciences LLC and Pioneer Hi-Bred International, Inc. request (00-136-01p) seeking a determina-tion of non-regulated status for Bt Cry1F insect resistant, glufosinate tolerant corn line 1507. United States Department of Agriculture, Animal and Plant Health Inspection Service, Washington D.C. http://www.aphis.usda.gov/brs/aphisdocs2/00_13601p_com.pdf
USDA APHIS (2002a). Application for an extension of the determination of nonregulated status for glufosinate-tolerant canola transformation (97-205-01p): Topas 19/2 (01-206-02) (HCN92). United States Department of Agriculture, An-imal and Plant Health Inspection Service, Washington D.C. http://www.aphis.usda.gov/brs/aphisdocs/01_20602p.pdf
USDA APHIS (2002b). Extension of determination of nonregulated status for canola genetically engineered for glufosinate herbicide tolerance (HCN92). Unit-ed States Department of Agriculture, Animal and Plant Health Inspection Serv-ice, Washington D.C. http://www.aphis.usda.gov/brs/aphisdocs2/01_20602p_com.pdf
USDA APHIS (2002c). Aventis CropScience USA LP petition for the determina-tion of nonregulated status for glufosinate-tolerant cotton transformation event, 02-042-01p (LLcotton25). United States Department of Agriculture, Animal and Plant Health Inspection Service, Washington D.C. http://www.aphis.usda.gov/brs/aphisdocs/02_04201p.pdf
USDA APHIS (2003a). Petition for determination of nonregulated status: B.t. Cry1F insect-resistant cotton event 281-24-236. United States Department of Agriculture, Animal and Plant Health Inspection Service, Washington D.C. http://www.aphis.usda.gov/brs/aphisdocs/03_03601p.pdf
USDA APHIS (2003b). Petition for determination of nonregulated status: B.t. Cry1Ac insect-resistant cotton event 3006-210-23. United States Department of Agriculture, Animal and Plant Health Inspection Service, Washington D.C. http://www.aphis.usda.gov/brs/aphisdocs/03_03602p.pdf
USDA APHIS (2003c). Environmental assessment and finding of no sig-nificant impact: Aventis CropScience USA LP petition for the determination of nonregulated status for glufosinate-tolerant cotton transformation event, 02-042-01p(LLcotton25). United States Department of Agriculture, Animal and Plant Health Inspection Service, Washington D.C. http://www.aphis.usda.gov/brs/aphisdocs2/02_04201p_com.pdf
USDA APHIS (2004a). Environmental assessment and finding of no sig-nificant impact: petition for determination of nonregulated status: B.t. Cry1F insect-resistant cotton event 281-24-236 and Cry1Ac insect-resistant cotton event 3006-210-23. United States Department of Agriculture, Animal and Plant Health Inspection Service, Washington D.C. http://www.aphis.usda.gov/brs/aphisdocs2/03_03602p_com.pdf
USDA APHISA (2004b). Application for an extension of the determination of nonregulated status for B.t. Cry1F insect-resistant, glufosinate tolerant maize (00-136-01p): maize line 6275. United States Department of Agriculture, Ani-mal and Plant Health Inspection Service, Washington D.C. http://www.aphis.usda.gov/brs/aphisdocs/03_18101p.pdf
13
USDA APHISA (2004c). Environmental assessment and finding of no signifi-cant impact: approval of Mycogen Seeds c/o Dow AgroSciences request (03-181-01p) seeking extension of the determination of nonregulated status for Bt Cry1F insect-resistant, glufosinate tolerant corn line 6275. United States Depart-ment of Agriculture, Animal and Plant Health Inspection Service, Washington D.C. http://www.aphis.usda.gov/brs/aphisdocs2/03_18101p_com.pdf
USDA APHISA (2004d). Application for the determination of nonregu-lated status for B.t. Cry34/35Ab1 insect-resistant, glufosinate tolerant corn: corn line 59122. United States Department of Agriculture, Animal and Plant Health Inspection Service, Washington D.C. http://www.aphis.usda.gov/brs/aphisdocs/03_35301p.pdf
USDA APHISA (2005). Environmental assessment and finding of no sig-nificant impact: request 03-353-01p seeking a determination of non-regulated status for B.t. Cry34Ab1/35Ab1 insect-resistant, glufosinate tolerant corn line 59122-7. United States Department of Agriculture, Animal and Plant Health Inspection Service, Washington D.C. http://www.aphis.usda.gov/brs/aphisdocs2/03_35301p_com.pdf
USDA APHIS (2006a). Application for an Extension of the Determination of Nonregulated Status for Glufosinate-Tolerant Rice (98-329-01p): Transforma-tion Event LLRICE601. United States Department of Agriculture, Animal and Plant Health Inspection Service, Washington D.C. http://www.aphis.usda.gov/brs/aphisdocs/06_23401p.pdf
USDA APHIS (2006b). Environmental assessment and finding of no significant impact: extension of nonregulated status to rice line LLRice601. United States Department of Agriculture, Animal and Plant Health Inspection Service, Wash-ington D.C. http://www.aphis.usda.gov/brs/aphisdocs2/06_23401p_com.pdf
USEPA (2001). Biopesticide registration action document: Bacillus thuring-iensis Cry1F corn. United States Environmental Protection Agency, Washing-ton D.C. http://www.epa.gov/pesticides/biopesticides/ingredients/tech_docs/brad_006481.pdf
USEPA (2005). Bacillus thuringiensis Cry1F/Cry1Ac construct 281/3006 insec-ticidal crystal protein as expressed in cotton (006445 & 006481) fact sheet. Unit-ed States Environmental Protection Agency, Washington D.C. http://www.epa.gov/oppbppd1/biopesticides/ingredients/factsheets/factsheet_006445-6481.htm
USEPA (2009a). Bacillus thuringiensis Cry1Ab Delta-Endotoxin Protein and the Genetic Material Necessary for Its Production (via Elements of Vector pZO1502) in Event Bt11 Corn (OECD Unique Identifier: SYN-BT011-1)(006444) & Bacil-lus thuringiensis Vip3Aa20 Insecticidal Protein and the Genetic Material Neces-sary for Its Production (via Elements of Vector pNOV1300) in Event MIR162 Maize (OECD Unique Identifier: SYN-IR162-4)(006599) Fact Sheet. United States Environmental Protection Agency, Washington D.C. http://www.epa.gov/opp00001/biopesticides/ingredients/factsheets/factsheet_006599-006444.html
USEPA (2009b). Pesticide Fact Sheet (MON 89034 x TC1507 x MON 88017 x DAS-59122-7). United States Environmental Protection Agency, Washington D.C. http://www.epa.gov/oppbppd1/biopesticides/pips/smartstax-factsheet.pdf
annex I: summary oF Pat ProteIn exPressIon data
The tables that follow present summary data from peer-reviewed publications and regulatory submissions. Additional information on collection and sampling methodologies can be found in the ref-erenced sources.
Table I.1. Quantities of PAT in Beta vulgaris event T120-7 as detected by ELISA (USDA APHIS 1998b).
Plant Matrix1 % Protein2 ng PAT/g protein3,4
Roots 6.8 137
Tops (above ground) 15.0 966
Pulp (dried) 9.7 N.D.
Molasses 9.9 N.D.
1 Values reported are mean values from all sites2 Literature values (see USDA APHIS 1998b for citation).3 Two extracts from each sample (18 tops; 18 roots from 6 field sites) were analyzed in triplicate.4 Limit of Detection = 2 ng/g root; 1.6 ng/g sugar, pulp; 0.4 ng/g molasses .5 N.D. = Not Detected.
Table I.2. PAT contents in seed samples for Brassica napus event Topas 19/2 (HCN10 and HCN92) as detected by ELISA (USDA APHIS 2002a).
Sample ID PAT / Sample (ng/g)
Excel 1996 (control) N.D. 1
HCN92 295
HCN92 295
HCN10 189
HCN10 202
1 N.D. < Limit of Quantification (0.40 ng/mL).
Table I.3. PAT expression in seed and leaf samples from Brassica napus lines Topas 19/2, T45, and Topas 19/2 x T45 as detected by ELISA (USDA APHIS 2002a).
Sample ID Line/Treatment Event PAT/Sample (ng/g)
Total Protein (mg/g)
PAT/Protein
(%)Control Topas 19/2 ND 9.51
Plot 2 SW9782179 Topas 19/2 248 2.14 0.012
Plot 7 SW9782179 Topas 19/2 263 3.04 0.009
Plot 8 SW9782179 Topas 19/2 309 2.24 0.014
Plot 10 SW9782179 Topas 19/2 379 2.61 0.015
Plot 3 SW9782180 T45 555 2.68 0.021
Plot 5 SW9782180 T45 743 2.41 0.031
Plot 6 SW9782180 T45 717 2.17 0.033
Plot 1 SW9782213 Topas 19/2//T45 754 2.01 0.038
Plot 4 SW9782213 Topas 19/2//T45 906 2.00 0.045
Plot 9 SW9782213 Topas 19/2//T45 932 3.37 0.028
Plot 11 SW9782213 Topas 19/2//T45 944 3.12 0.030
Seed – UN Untreated Topas 19/2//T45 563 54.3 0.00104
Seed-TR Treated1 Topas 19/2//T45 669 59.4 0.00113
Tmeal-UN Untreated Topas 19/2//T45 ND 85.9 ND
Tmeal-TR Treated1 Topas 19/2//T45 ND 76.1 ND
RBD oil - UN Untreated Topas 19/2//T45 ND ND ND
RBD oil - TR Treated1 Topas 19/2//T45 ND ND ND
1 Treated with glufosinate.
14
Table I.4. PAT expression in seeds and leaves of Brassica napus lines RF3 and MS8 determined by enzyme activity (OGTR 2003).
GM Canola Line Seed µg PAT/mg Total Protein
Leaf µg PAT/mg Total Protein
RF3 0.10 1.33
MS8 0.04 0.51
RF1 Not tested 1.45
RF2 Not tested 0.7
MS1 Not tested 0.9
Table I.5. PAT expression in seeds of GM Brassica napus lines determined by ELISA (OGTR 2003).
GM Canola Line µg PAT/g Seed µg PAT/mg Total Protein
RF3 0.69 0.012
MS8 0.07 0.002
RF3xMS8 0.34 0.013
RF1 0.50 0.015
RF2 0.42 0.012
MS1 0.07 0.002
MS1xRF1 0.20 0.006
MS1xRF2 0.35 0.007
Table I.6. PAT expression in seeds of GM Brassica napus lines deter-mined by ELISA (OGTR 2003).
GM Canola Line Seed µg PAT/g Total Seed
Leaf µg PAT/mg Fresh Weight
T45 0.561 0.348
Topas 0.47 0.0843
Table I.7. Summary of mRNA expression analysis for bar in Brassica napus RF2.
Total RNA pg bar mRNA/µg Total RNA(Range of Detected Values)
Leaves 0.8-1.6
Flower Buds 0.1-0.2
Seed ND1
Pollen ND
1 N.D. = Not Detected (<2pg/ µg total RNA).
Table I.8. Summary of mRNA expression analysis for bar in Brassica napus MS8 (USDA APHIS 1998l)1.
Total RNA Transgene Expression (pg/µg Total RNA)2
Leaf A 0.03
Leaf B 0.22
Flower buds 2mm A 0.14
Flower buds 2mm B 0.11
Flower buds 3mm A 0.19
Flower buds 3mm B 0.03
Dry seed ND3
1 Data shown for two plants (A and B) with single samples of each tissue.2 pGembar/SP6 plasmid used for preparation of RNA probe.3 ND = Not Detected. Limit of Detection = 0.1pg/g total RNA.
Table I.9. Summary of mRNA expression analysis for bar in Brassica napus RF3 (USDA APHIS 1998l).
Total RNA Transgene Expression (pg/µg Total RNA)1
Leaf A 1.1
Leaf B 0.2
Flower buds 2mm A 0.46
Flower buds 2mm B 0.52
Flower buds 3mm A 0.38
Flower buds 3mm B 0.34
Dry seed ND2
Pollen ND2
1 Data shown for two plants (A and B) with single samples of each tissue.2 ND = Not Detected. Limit of Detection = 0.05pg/g total RNA.
Table I.10. Summary of PAT expression in seed of Brassica napus lines T45 (HCN28) and HCN 92 (method not reported) (CFIA 1996b).
Sample µg/mg Sample (Reported Range)
T45 Seed 95-245
HCN 92 Seed 150-223
Table I.11. Summary of PAT expression in Brassica napus line T45 (HCN28) using Northern blot and ELISA (EFSA 2008).
Sample Northern Blot1 ELISA
Leaves + NA2
Stems + NA
Roots + NA
Seeds - 930 ng/g dry weight
1 Results are reported as presence (+) or absence (-) of detectable mRNA.2 NA = Not Available.
Table I.12. Summary of PAT content of seed from Brassica rapa line HCR-1 (method not reported) (CFIA 1998c).
Tissue Mean (ng/g) Range (ng/g)
Seed 107 84-132
15
Table I.13. Summary of PAT protein in seeds of Glyxine max lines AS2704-12 and A5547-127 as detected by ELISA (FSANZ 2004a).
Sample Year Treatment1 PAT/Sample (ng/g)
Crude Protein (%)
PAT Protein as % of Crude Protein
AS2704-12 1997 NA2 573 37-45 0.00016
A5547-127 1998 NA 10800 37-45 0.00292
AS2704-12 1999 sprayed 879 (264)3 NA 0.000227
AS2704-12 1999 unsprayed 862 (272) NA 0.000227
A5547-127 1999 sprayed 10100 (816) NA 0.00285
A5547-127 1999 unsprayed 9971 (940) NA 0.00283
AS2704-12 NA sprayed 2183 38.9 0.00050
AS2704-12 NA unsprayed 1948 38.5 0.00056
A5547-127 NA sprayed 17471 36.5 0.0048
A5547-127 NA unsprayed 20202 35.8 0.0056
1 sprayed = treated with glufosinate; unsprayed = not treated with glufosinate.2 NA = Not Available.3 Mean (Standard Deviation).
Table I.14. Summary of PAT protein detected in Glycine max line A2704-12 using ELISA (Japan BCH 1999b).
Tissue Mean PAT (µg/g Fresh Weight)
± Standard Deviation
Crude Protein (% of Fresh Weight)
PAT Protein (% of Crude Protein)
Root 2.23 ± 1.29 1.95 0.011
Stem 7.63 ± 2.20 3.58 0.021
Leaf 14.5 ± 2.4 5.96 0.024
Table I.15. Summary of PAT protein detected in seeds of Glycine max line A2704-12 using ELISA (Japan BCH 1999b).
Sample PAT (ng/g Sample) Mean ± Standard Deviation
Crude Protein Content (%)
PAT/Crude Protein (%)
1 1057 NA1 NA
2 573 NA NA
3 862 ± 268 38.03 0.000227
4 2138 ± 33 43.5 0.00049
1 NA = Not Available.
Table I.18. Summary of PAT content in leaves of Glycine max line A5547-127 as detected by ELISA (USDA APHIS 1998d)1.
Sample mg TEP2/g Sample
µg PAT/ g Sample
% PAT/ TEP
% PAT/ Fresh Weight g/g
A5547-127 4.6 1.72 0.037 1.72 x 10-4
1 Values are the average from two replicate extractions from two samples of 10 day old seedling leaves.2 TEP = Total Extractable Protein.
Table I.19. Summary of PAT content in leaves of Glycine max line GU262 as detected by ELISA (USDA APHIS 1998f )1.
Sample mg TEP2/g Sample
µg PAT/ g Sample
% PAT/ TEP
% PAT/ Fresh Weight g/g
GU262 4.7 3.03 0.064 3.03 x 10-4
1 Values are the average from two replicate extractions from two samples of 10 day old seedling leaves.2 TEP = Total Extractable Protein.
Table I.20. Summary of PAT protein in Glycine max lines W62 and W98 as detected by enzymatic activity (USDA APHIS 1996a)1.
Tissue Site and Year Plant µg PAT/g Sample2
Fodder (whole plant) Arkansas 1993 W62 10.8 (6.3-15.3)
Iowa 1993 W98 0.75 (0.65-1.0)
Illinois 1993 W98 10.9 (9.1-12.7)
Seed Arkansas 1993 W62 217.0 (147.1-267.3)
Iowa 1993 W98 27.1 (15.0-39.2)
Illinois 1993 W98 38.3 (23.5-60.9)
1 Values are the average from two replicate extractions from two samples of 10 day old seedling leaves.2 Mean (range of observed values).
Table I.16. Summary of PAT protein detected in Glycine max line A5547-127 using ELISA (Japan BCH 2006f ).
Tissue Mean PAT (µg/g Fresh Weight)
± Standard Deviation
Crude Protein (% of Fresh Weight)
PAT Protein (% of Crude Protein)
Root 3.73. ± 0.98 2.15 0.017
Stem 11.5 ± 1.8 3.62 0.032
Leaf 19.0 ± 5.0 6.70 0.028
Table I.17. Summary of PAT protein detected in seeds of Glycine max line A5547-127 using ELISA (Japan BCH 2006f ).
Sample PAT (ng/g Sample) Mean ± Standard Deviation
Crude Protein Content (%)
PAT/Crude Protein (%)
1 6341 NA1 NA
2 10800 ± 1210 NA NA
3 9971 ± 846 35.26 0.00282
4 20202 ± 359 40.4 0.0050
1 NA = Not Available.
16
Table I.22. Summary of PAT protein expression in Gossypium hirsutum even 3006-210-23 as determined by ELISA (USDA 2004a).
Tissue PAT (ng/mg Dry Weight)
Mean1 Standard Deviation
Minimum-Maximum Range
Young Leaf (3-6 week) ND4 NA3 0.18-0.67
Terminal Leaf ND NA ND-0.12
Flower ND NA ND-ND
Square ND NA ND-0.08
Boll (early) ND NA ND-0.08
Whole Plant (seedling)
ND NA ND-0.09
Whole Plant (pollination)
ND NA ND-0.14
Whole Plant (defoliation)
0.11 0.05 ND-0.20
Root (seedling) ND NA ND-0.07
Root (pollination) ND NA ND-ND
Root (defoliation) ND NA ND-ND
Pollen5 ND NA ND-ND
Nectar5 ND NA ND
Seed5 0.062 0.06 ND-0.232
1 Calculated from samples at six locations.2 Value below the limit of quantification of the method.3 NA = Not Applicable.4 ND = absorbance of the sample was lower than the absorbance of the lowest standard.5 Results are given relative to fresh weight.
Table I.23. Summary of PAT protein expression in Gossypium hirsutum even 3006-210-23 as determined by ELISA (USDA 20036).
Tissue Mean Protein Expression (ng/mg dry weight)
Young Leaf (3-6 week) 0.43
Terminal Leaf 0.23
Flower 0.35
Square 0.52
Boll (early) 0.27
Whole Plant (seedling) 0.35
Whole Plant (pollination) 0.30
Whole Plant (defoliation) 0.34
Root (seedling) 0.062
Root (pollination) ND
Root (defoliation) 0.052
Pollen1 0.052
Nectar1 ND
Seed1 0.54
1 Results reported as ng/mg fresh weight.2 Calculated concentration is less than the limit of quantification of the method.
Table I.24. Summary of PAT expression in Gossypium hirsutum line LLCotton25 measured by ELISA (CFIA 2004).
Tissue PAT Protein (µg/g Fresh Weight)
Root 7.97
Leaf 52.9
Stem 36.8
Fuzzy Seed 69.9
Cleaned Seed 127
Pollen 19.2
Table I.25. Summary of PAT protein content in Gossypium hirsutum line LLCotton25 as measured by ELISA (OGTR 2006).
Tissue PAT (µg/g Fresh Weight)
PAT as % of Crude Protein
Average TEP2 (mg/g
Fresh Weight ±
SD)
Average PAT Content as % of TEPRange Average
Roots 5.63-10.1 7.97 ± 1.86 0.08 2.26 ± 0.22 0.35
Stems 34.3-45.5 36.8 ± 6.7 0.23 4.99 ± 0.92 0.74
Leaves 45.1-57.3 52.9 ± 6.0 0.19 7.13 ± 0.79 0.74
Pollen (frozen) 4.44-13.0 8.23 ± 3.20 NA1 146 ± 8 0.006
Pollen (fresh) 0.11-170 19.3 ± 39.2 NA 107 ± 21 0.018
1 Not Applicable.2 TEP = Total Extractable Protein.
Table I.21. Summary of PAT protein expression in Gossypium hirsutum event 281-24-236 as determined by ELISA (USDA 2003a).
Tissue PAT (ng/mg Dry Weight)
Mean1 Standard Deviation
Minimum-Maximum Range
Young Leaf (3-6 week) 0.43 0.12 0.18-0.67
Terminal Leaf 0.21 0.12 ND4-0.38
Flower 0.29 0.11 0.072-0.44
Square 0.51 0.15 0.062-0.79
Boll (early) 0.22 0.09 0.082-0.48
Whole Plant (seedling)
0.31 0.07 0.21-0.46
Whole Plant (pollination)
0.23 0.07 0.092-0.33
Whole Plant (defoliation)
0.19 0.13 ND-0.46
Root (seedling) 0.072 0.05 ND-0.12
Root (pollination) ND NA3 ND-0.11
Root (defoliation) ND NA ND-0.11
Pollen5 0.092 0.15 ND-0.45
Nectar5 ND NA ND-ND
Seed5 0.47 0.17 0.232-1.02
1 Calculated from samples at six locations.2 Value below the limit of quantification of the method.3 NA = Not Applicable.4 ND = absorbance of the sample was lower than the absorbance of the lowest standard.5 Results are given relative to fresh weight.
17
Table I.26. Summary of PAT protein content in leaves of Gossypium hirsutum line LLCotton25 as measured by ELISA (OGTR 2006).
Sample PAT Protein (µg/g Fresh Weight ± SD)
2-4 leaf 4-5 leaf Early Bloom Full Bloom
Non GM Control
ND1 ND ND ND
Sprayed Once2 NA 85.0 ± 15.6 98.3 ± 16.8 92.6 ± 15.1
Sprayed Twice NA NA NA 92.6 ± 20.3
Unsprayed 57.7 ± 5.3 74.0 ± 12.3 90.2± 14.4 75.1 ± 25.6
1 ND = not detected; NA = not applicable.2 Sprayed with Liberty Link.
Table I.27. Summary of PAT protein content RACs of Gossypium hirsu-tum line LLCotton25 as measured by ELISA (OGTR 2006).
Sample Average PAT Protein (µg/mg Fresh Weight) ± SD
Average Protein Content (as % of Crude Protein)
LL Sprayed Unsprayed LL Sprayed Unsprayed
Cleaned Seed 127 ± 18 113 ± 24 NA1 NA
Lint Coat 1.5 ± 0.45 0.92 ± 0.50 NA NA
Fuzzy Seed 69.9 ± 6.0 63.0 ± 10.3 0.030 0.027
Lint 0.78 ± 0.63 0.50 ± 0.42 0.003 0.003
1 NA = Not Applicable.
Table I.28. Summary of PAT protein in Gossypium hirsutum line LLCot-ton25 as measured by ELISA (USDA APHIS 2002c).
Sample PAT Protein (µg/g Fresh Weight)
± Standard Deviation
PAT Content (% of Crude Protein)
Liberty Herbicide
Conventional Herbicide
Liberty Herbicide
Conventional Herbicide
Cleaned Seed 127 ± 18 113 ± 24 NA NA
Lint Coat 1.15 ± 0.45 0.92 ± 0.50 NA NA
Fuzzy Seed 69.9 ± 6.0 63.0 ± 10.3 0.030 0.027
Lint 0.78 ± 0.63 0.50 ± 0.42 0.003 0.003
Table I.29. Summary of PAT protein content in leaves of Gossypium hirsutum line LLCotton25x15985 as measured by ELISA (Japan BCH 2006a).
Sample Protein Content(Mean ± Standard Deviation)
(µg/g of Leaf )LLCotton25 x 15985 60.9 ± 8.71
LLCotton25 65.9 ± 10.61
1 The difference in expression between the stacked line and the single event is not significant.
Table I.30. Summary of PAT protein content in Oryza sativa line LL-Rice62 as measured by ELISA (CFIA 2006b).
Tissue Mean Protein Content µg/g Fresh Weight
Grain 12.1
Straw 75.3
Rice Hulls 1.56
Roots 12.7
Stems 30.9
Leaves 84.7
Table I.31. Summary of PAT protein content in Oryza sativa line LL-RICE62 as measured by ELISA (FSANZ 2008).
Tissue Average PAT Content (µg/g ± SD)
Crude Protein (% w/w)
PAT Protein (% of Crude
Protein)Grain (Year 1) 12.1 ± 0.6 7.19 0.017
Straw (Year 1) 75.3 ± 4.4 2.38 0.316
Grain (Year 2) 10.6 ± 1.3 7.41 0.014
Table I.32. Summary of PAT protein content in grain of Oryza sativa lines LLRICE06 and LLRICE62 as detected by ELISA (USDA APHIS 1998h).
Plant mg TEP1/g Sample
µgPAT/g Sample
% PAT/TEP % PAT/Fresh Weight (g/g)
LLRICE06 1.89 ± 0.49 0.419 ± 0.04 0.02 0.00005
LLRICE62 2.54 ± 0.09 12.4 ± 2.4 0.63 0.00124
1 TEP = Total Extractable Protein
Table I.33. Summary of PAT protein content in seed of Oryza sativa line LLRICE601 (method not reported) (USDA APHIS 2006a).
Plant Protein Content (ng/g Fresh Weight)
% of Crude Rice Protein
LLRICE601 120 0.00034
18
Table I.36. Expression of PAT protein in Zea mays line B16 (method not reported) (CFIA 1996d).
Tissue PAT Protein Detected
Leaves 1.0-4.6 mg/g protein
Roots +1
Stalk +
Tassel +
Cob +
Husk +
Kernels -2
Silk -
Pollen -
1 + = Protein detected but quantity not reported.2 - = Protein not detected.
Table I.37. Expression of PAT protein in Zea mays line B16 determined by Western blot (USDA APHIS 1995b).
Tissue PAT Concentration (ng/µg Total Protein)
PAT Concentration (ng/mg Fresh Weight)
Coleoptile (6 days) 1.8 13.8
Leaf (24 days) 1.0 55.6
Leaf (44 days) 2.8 ± 0.1 166.0 ± 24.2
Leaf (93 days) 2.1 106.1
F¬2 ovule (0 days pp) 0.8 5.5
Immature F¬2¬ seed (16 days pp)
0.3 3.4
Immature F¬2 seed (45 days pp)
0.3 5.7
Hybrid seed (F1) 0.2 1.9
Root (24 days) 0.06 4.2
Root (44days) 1.3 8.1
Prop root (49 days) 1.9 ±0.3 19.8 ± 2.4
Cob (56 days) 2.2 67.2
Husk (56 days) 1.1 2.5
Silk ND1 4.2
Stalk (24 days) 2.0
Stalk (77 days) 4.6 ± 0.4 15.7
Immature tassel (49 days) 2.0 11.2 ± 2.7
Pollen ND 30.8
Silage ND
1 ND = Not detected. <0.05 ng/mg in silk; <0.08 ng/mg in pollen and silage.
Table I.38. Mean level of expression of the PAT protein in Zea mays line Bt11 (X4334-CBR and X4743) using ELISA (ANZFA 2001a).
Mean (µg/g Fresh Weight)
Leaf Kernel Husk Stalk
X4334-CBR 0.0386 ± 0.0029 lod1 lod lod
X4734-CBR 0.0494 ± 0.005 lod nd2 nd
Control (NK4242) lod lod lod lod
1 lod (limit of detection) for the procedure is 1ng PAT/mL extract. These values are considered not above background.2 nd = No data.
Table I.34. PAT protein expression in Zea mays line Bt-176 and hybrid plants as determined by ELISA (USDA APHIS 1995a)1.
PAT Expressionng/g Fresh Weight
Seedling Anthesis Seed Maturity
Senescence
Leaves Bt176 <2004 <200 <200 ND
176 x 5542 <200 ND ND ND
176 x 5643 <200 ND <200 <200
Whole Plant Bt176 ND <200 <200 <200
176 x 554 <200 ND <200 <200
176 x 564 <200 ND ND <200
Kernels Bt176 ND ND
176 x 554 ND ND
176 x 564 ND ND
Pollen Bt176 ND
176 x 554 NA
176 x 564 ND
Roots Bt176 ND4 <100 <100 NA
176 x 554 ND <100 <100 NA
176 x 564 ND ND <100 NA
Pith Bt176 NA5 <200 <200 NA
176 x 554 NA ND <200 NA
176 x 564 NA NA ND NA
1 Blank cell indicate no developmental relevance.2 176 x 554 = hybrid progeny of CG00526-176 and untransformed CG00554 and is hemizygous for the introduced genes.3 176 x 564 = hybrid progeny of CG00526-176 and untransformed CG00564 and is hemizygous for the introduced genes.4 When trace amounts were detectable but not quantifiable, results are shown as < lower limit of quantification.5 ND = Not Detected.6 NA = Not Analyzed.
Table I.35. Expression of PAT protein in leaf tissue of male sterile Zea mays lines 676, 678, and 680 as determined with ELISA (USDA APHIS 1997d).
Male Sterile Corn Line PAT Concentration µg/g Total Protein
676 601-617
678 204-278
680 <201
1 Below the limit of quantification (20µg/g).
19
Table I.40. Mean expression of PAT protein in Zea mays line CBH-351 using ELISA (USDA APHIS 1997e)1.
Tissue Stage 1 Stage 2 Stage 3 Stage 4
Total Protein (mg/g)
PAT Protein (µg/g)
Total Protein (mg/g)
PAT Protein µg/g
Total Protein (mg/g)
PAT Protein µg/g
Total Protein (mg/g)
PAT Protein µg/g
Whole Plant 12.5 ± 4.6 189.7 ± 23.5 7.8 ± 1.9 105.7 ± 16.7 5.7 ± 1.4 44.4 ± 5.4 2.9 ± 1.6 6.8 ± 1.6
Leaf 3.1 ± 1.3 45.4 ± 9.7 0.6 ± 0.1 14.0 ± 0.1 1.5 ± 0.1 2.3 ± 0.2 1.8 ± 0.2 0.0 ± 0.0
Root 0.7 ± 0.1 39.1 ± 2.7 25.8 ± 19.2 4.4 ± 0.8 0.3 ± 0.2 0.6 ± 0.5 0.6 2.2 ± 2.8
Stalk NA2 NA 2.8 ± 1.2 0.5 ± 0.5 0.0 ± 0.0 0.5 ± 0.2 ± 0.2 0.0 → 0.1
Tassel NA NA 175.0 ± 100.4 4.2 ± 1.2 NA NA NA NA
Kernel NA NA NA NA NA NA 6.7 ± 1.1 17.8 ± 5.0
1 Values are expressed as mean ± standard deviation. All values are relative to dry weight of samples.2 NA = Not Applicable.
Table I.39. Mean level of expression of the PAT protein in Zea mays line Bt11 (method not reported) (CFIA 1996c).
Mean (µg/g fresh weight)
Leaf Tassel Silk Roots Kernel Pollen
BT11 0.049 0.027 0.005 ND1 ND ND
1 ND = Not Detected.
Table I.41. Mean level of PAT protein expression in hybrids derived from Zea mays line CBH-351 using ELISA (USDA APHIS 1997e)1.
Hybrid Stage 1 Stage 2
Total Protein (mg/g)
PAT Protein µg/g
Total Protein (mg/g)
PAT Protein µg/g
Hybrid A 19.1 ± 8.2 364.0 ± 85.1 4.9 ± 2.0 40.7 ± 16.7
Hybrid B 18.3 ± 1.1 291.0 ± 4.2 7.2 ± 2.8 77.1 ± 12.9
Hybrid C 11.6 ± 6.3 227.0 ± 33.4 6.8 ± 1.3 100.2 ± 24.9
Hybrid D 9.1 ± 3.3 176.3 ± 63.0 4.1 ± 1.4 88.3 ± 23.4
1 Values are expressed as mean ± standard deviation. All values are relative to dry weight of samples.
Table I.42. Mean expression of PAT protein in Zea mays line DAS-06275-8 at six field trial sites in the U.S. and Canada as measured by ELISA (CFIA 2006a).
Leaf Root Stalk Grain Pollen Forage
PAT Protein Expression (ng/mg Dry Weight)
129.2-224.21 61.1-70.01 103.3 8.94 0.73 106.9
1 Range represents means measured across multiple growth stages.
Table I.43. Mean expression of PAT protein in Zea mays line DAS-06275-8 in grain (method not reported) (Health Canada 2006).
U.S. Chile
PAT Protein Expression (ng/mg Dry Weight)
5.94 23
Table I.44. Summary of mean expression of BAR (PAT)protein in a Zea mays hybrid derived from DAS-06275-8 measured by ELISA (USDA APHIS 2004b).
Growth Stage Tissue Mean ± Standard Deviation
(ng/mg Dry Weight)
Minimum-Maximum Range
(ng/mg Dry Weight)V9 Leaf 323 ± 91.0 0-538
Root 112 ± 35.3 0-170
Whole plant 5 ± 3.50 1-11
R1 Leaf 674 ± 98.1 539-935
Root 253 ± 162 61-673
Whole plant 72 ± 32.9 35-108
R4 Pollen 0 ± 0.766 0-4.07
Stalk 282 ± 68.5 177-475
Leaf 682 ± 254 451-1584
Maturity Root 223 ± 105 85-511
Forage 7 ± 7.05 1-19
Grain 23 ± 6.33 13-33
Senescence Leaf 0 ± 0.461 0-1
Root 41 ± 49.5 0-148
Whole plant 18 ± 5.27 9-23
Table I.45. Summary of PAT protein expression in Zea mays line DAS-59122-7 at multiple sites in the U.S. and Canada measured by ELISA (CFIA 2005).
Leaf Root Stalk Grain Pollen Forage
PAT Protein Expression (ng/mg Dry Weight)
0.25-11.41 0.18-0.421 0.38 0.1 LOD2 2.4
1 Range represents means measured across multiple growth stages.2 LOD = Below the Limit of Detection (<0.30 ng/mg dry weight).
20
Table I.46. Summary of expression levels of PAT protein in Zea mays line DAS-59122-7 as measured by ELISA (USDA APHIS 2004d).
Growth Stage Tissue Mean ± Standard Deviation
(ng/mg Dry Weight)
Minimum-Maximum Range
(ng/mg Dry Weight)V9 Leaf 11.1 ± 3.68 5.61-19.2
Root 0.47 ± 0.15 0.27-0.87
Whole plant 0.18 ± 0.13 0-0.40
R1 Leaf 11.2 ± 3.49 6.36-18.2
Root 0 ± 0 0-0
Whole plant 0.13 ± 0.03 0.07-0.20
R4 Pollen 0.27 ± 0.12 0.11-0.62
Stalk 0.13 ± 0.23 0-0.58
Leaf 8.13 ± 3.02 0-14.2
Maturity Root 0.09 ± 0.12 0-0.34
Forage 0 ± 0 0-0
Grain 0 ± 0 0-0
Senescence Leaf 0.38 ± 0.46 0-1.33
Root 0.08 ± 0.11 0-0.46
Whole plant 0 ± 0 0-0
Table I.47. Summary of expression levels of PAT protein in grain of Zea mays hybrids with line 59122 as detected by ELISA (EFSA 2008b/EFSA 2009a).
59122 x 1507 x NK603
59122 X NK603 59122
Mean Range Mean Range Mean Range
PAT (ng/mg Dry Weight)
2.1 1.5-3.1 0.18 0-0.46 0.09 0-0.18
Table I.48. Summary of expression levels of PAT protein in Zea mays line DBT418 using Western blot (USDA 1996d).
Tissue Genotype Mean Protein Level (µg/g Dry Weight)
V6-V7 Pollen Shed Dough Harvest
Mean n ; SE Mean n ; SE Mean n ; SE Mean n ; SE
Leaf Inbred1 351.1 7; 52.91 522.0 6; 59.04 NA NA 60.86 6; 12.46
Hemizygous hybrid2 276.3 8; 25.51 501.8 8; 34.75 NA NA 180.5 8; 24.68
Homozygous hybrid3 554.9 2; 136.03 1099.4 3; 76.29 NA NA 213.6 4; 61.92
Stalk Inbred NA NA 75.8 8; 12.24 NA NA 95.2 6; 16.86
Hemizygous hybrid NA NA 60.0 8; 11.98 NA NA 64.4 8; 8.23
Homozygous hybrid NA NA 77.0 4; 11.66 NA NA 136.3 2; 12.74
Root Ball Inbred 95.1 7; 16.91 54.1 8; 9.15 NA NA 24.5 7; 3.71
Hemizygous hybrid 59.4 8; 3.53 27.5 8; 6.25 NA NA 21.3 8; 2.23
Homozygous hybrid 88.1 4; 21.45 69.5 4; 23.58 NA NA 28.8 3; 7.37
Kernel Inbred NA NA NA NA NA NA 6.0 6; 1.88
Hemizygous hybrid NA NA NA NA NA NA 3.1 8; 0.35
Homozygous hybrid NA NA NA NA NA NA 4.9 4; 0.63
Silk Inbred NA NA 128.2 8; 17.21 NA NA NA NA
Hemizygous hybrid NA NA 29.1 8; 2.97 NA NA NA NA
Homozygous hybrid NA NA 133.3 2; 60.01 NA NA NA NA
Pollen Hemizygous hybrid4 NA NA BLD5 8; NA NA NA NA NA
Hemizygous hybrid NA NA BLD 8; NA NA NA NA NA
Homozygous hybrid NA NA BLD 4; NA NA NA NA NA
Whole Plant Inbred NA NA 111.1 8; 16.50 190.5 8; 30.76 NA NA
Hemizygous hybrid NA NA 72.8 8; 5.88 39.5 7; 7.51 NA NA
Homozygous hybrid NA NA 119.5 4; 25.63 135.2 3; 10.42 NA NA
1 The S4 inbred line (AW/BC2/DBT418 S4) is an unfinished inbred.2 This hybrid (AW/BC2/DBT418.BS/BC1/DBT418) contains two integrated copies of DBT418 insertion.3 This hybrid (DK.DL (DBT418)) is a “finished hybrid” with one copy of DBT418 coming from an inbred parent line.4 An additional group of the Hemizygous hybrid (AW/BC2/DBT418.BS/BC1/DBT418) was substituted because insufficient pollen was available from the S4 hybrid.5 Below the Limit of Detection (12.10 µg/g dry weight).6 2 of 8 samples were below the limit of detection and not used to calculate the mean or standard error.
21
Table I.49. Summary of the quantification of bar mRNA transcripts in Zea mays line MS3 (method not reported) (CFIA 1996e).
Line Approximate mRNA (pg/µg total RNA)
Leaves Immature Kernels
Roots Dry Seeds2 Germinating Seeds2
MS3 0.05 0.05 ND1 ND ND
1 ND = Not Detected.2 A PAT enzyme activity assay did not detect any PAT in MS3 seeds.
Table I.50. Summary of the PAT protein content Zea mays line MS6 as detected by ELISA (USDA APHIS 1998k).
Tissue Mg TEP1/g Sample
µg PAT/g Sample % PAT/TEP
Grain 8.73 3.54 0.04
Forage 1.31 2.01 0.15
Fodder 1.26 2.15 0.17
1 TEP = Total Extractable Protein.
Table I.51. Summary of PAT protein levels in Zea mays hybrid and inbred lines derived from T25 as detected by ELISA (ANZFA 2001b).
Plant Mean Levels ± Standard Deviation (ng/mg Protein)
Kernel Silage Forage Fodder
Hybrid T25-1 ND2 14.82 ± 0.86 NA3 NA
Hybrid T25-2 ND 12.51 ± 1.38 NA NA
Hybrid T25-31 ND 14.81 ± 1.30 NA NA
Inbred T25 4.02 ± 0.62 119.24 ± 13.36 62.70 ± 40.07 79.91 ± 5.23
1 Plants were treated with phosphinothricin at the V8 stage.2 ND = Not Detected.3 NA = Data Not Available.
Table I.52. Summary of PAT protein quantities detected by ELISA in Zea mays lines T14 and T25 (USDA 1994b) 1.
Matrix % Protein ng PAT/µg Protein µg PAT/g Matrix % PAT in Matrix
T14 silage 0.19 13.03 36.97 3.70
T25 silage 0.05 13.54 6.62 0.67
T14 grain 1.59 0.008 0.115 0.0115
1 Two extracts from each sampel (2 each for silage, 6 for grain) were analyzed in triplicate. Means are reported from all field sites combined.
Table I.53. Summary of PAT protein expression in Zea mays line T25 and hybrid crosses with MON810 as detected by ELISA.
Tissue T25 X MON810 T25
Mean Min-Max Mean Min-Max
Leaves 33.3 17.1-54.5 33.9 11.9-64.6
Table I.54. Summary of PAT protein expression as determined by ELISA in Zea mays line TC1507(CFIA 2002b, EFSA 2005) 1.
Location of Cultivation
Tissue
Leaf Tassel Silk Roots Kernel Pollen
Canada <LOD2 <LOD <LOD <LOD <LOD NA
Chile <LOD <LOD <LOD <LOD <LOD NA
EU 42 pg/µg (<LOD-136.8
pg/µg)3
<LOD <LOD <LOD <LOD NA
United States <LOD <LOD <LOD <LOD <LOD <LOD (<LOD-38.0 pg/µg)
1 Data presented are from descriptive paragraphs describing different aspects of the same data set. These have been combined for simplicity.2 LOD = Limit of Detection (7.5 pg/µg total protein for samples from Canada, 20 pg/µg for all other locations.3 Mean (Range).
Table I.55. SSummary of PAT protein expression as determined by ELISA in Zea mays line TC1507 (USDA 2001c) 1.
Leaf Pollen Silk Stalk Whole Plant
Grain Senescent Whole Plant
<LOD2 (<LOD-40.8) <LOD <LOD <LOD <LOD <LOD <LOD
1 All values are pg/µg protein. Mean values are listed with observed ranges in parentheses.2 LOD = Limit of Detection (20 pg/µg total protein).
Table I.56. Summary of PAT protein expression in plants derived from Zea mays line TC1507 and DAS59122 as detected by ELISA (EFSA 2009b) 1.
1507 x5912 1507 59122
Grain Mean Range Mean Range Mean Range
0.15 0.09-0.27 <LOD2 <LOD 0.09 0-0.18
Forage 2.53 1.01-3.97 NA NA NA NA
1 Values are ng/mg dry weight.2 LOD = Limit of Detection.
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